Hydrogel Extrusion Speed Measurements for the Optimization of Bioprinting Parameters.

Three-dimensional (3D) bioprinting is the use of computer-controlled transfer processes for assembling bioinks (cell clusters or materials loaded with cells) into structures of prescribed 3D organization. The correct bioprinting parameters ensure a fast and accurate bioink deposition without exposing the cells to harsh conditions. This study seeks to optimize pneumatic extrusion-based bioprinting based on hydrogel flow rate and extrusion speed measurements. We measured the rate of the hydrogel flow through a cylindrical nozzle and used non-Newtonian hydrodynamics to fit the results. From the videos of free-hanging hydrogel strands delivered from a stationary print head, we inferred the extrusion speed, defined as the speed of advancement of newly formed strands. Then, we relied on volume conservation to evaluate the extrudate swell ratio. The theoretical analysis enabled us to compute the extrusion speed for pressures not tested experimentally as well as the printing speed needed to deposit hydrogel filaments of a given diameter. Finally, the proposed methodology was tested experimentally by analyzing the morphology of triple-layered square-grid hydrogel constructs printed at various applied pressures while the printing speeds matched the corresponding extrusion speeds. Taken together, the results of this study suggest that preliminary measurements and theoretical analyses can simplify the search for the optimal bioprinting parameters.

Assessment of RAS-RAF-MAPK Pathway Mutation Status in Healthy Skin, Benign Nevi, and Cutaneous Melanomas: Pilot Study Using Droplet Digital PCR.

In the present study, we employed the ddPCR and IHC techniques to assess the prevalence and roles of RAS and RAF mutations in a small batch of melanoma (n = 22), benign moles (n = 15), and normal skin samples (n = 15). Mutational screening revealed the coexistence of BRAF and NRAS mutations in melanomas and nevi and the occurrence of NRAS G12/G13 variants in healthy skin. All investigated nevi had driver mutations in the BRAF or NRAS genes and elevated p16 protein expression, indicating cell cycle arrest despite an increased mutational burden. BRAF V600 mutations were identified in 54% of melanomas, and NRAS G12/G13 mutations in 50%. The BRAF mutations were associated with the Breslow index (BI) (p = 0.029) and TIL infiltration (p = 0.027), whereas the NRAS mutations correlated with the BI (p = 0.01) and the mitotic index (p = 0.04). Here, we demonstrate that the "young" ddPCR technology is as effective as a CE-IVD marked real-time PCR method for detecting BRAF V600 hotspot mutations in tumor biopsies and recommend it for extended use in clinical settings. Moreover, ddPCR was able to detect low-frequency hotspot mutations, such as NRAS G12/G13, in our tissue specimens, which makes it a promising tool for investigating the mutational landscape of sun-damaged skin, benign nevi, and melanomas in more extensive clinical studies.

Diabetic neuropathy: A NRF2 disease?

The transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) has multifarious action with its target genes having redox-regulating functions and being involved in inflammation control, proteostasis, autophagy, and metabolic pathways. Therefore, the genes controlled by NRF2 are involved in the pathogenesis of myriad diseases, such as cardiovascular diseases, metabolic syndrome, neurodegenerative diseases, autoimmune disorders, and cancer. Amidst this large array of diseases, diabetic neuropathy (DN) occurs in half of patients diagnosed with diabetes and appears as an injury inflicted upon peripheral and autonomic nervous systems. As a complex effector factor, NRF2 has entered the spotlight during the search of new biomarkers and/or new therapy targets in DN. Due to the growing attention for NRF2 as a modulating factor in several diseases, including DN, this paper aims to update the recently discovered regulatory pathways of NRF2 in oxidative stress, inflammation and immunity. It presents the animal models that further facilitated the human studies in regard to NRF2 modulation and the possibilities of using NRF2 as DN biomarker and/or as target therapy.

Compendium of Safety Regulatory for Safe Applications of Aerogels.

An increasing number of aerogels as nanostructured highly porous materials are entering the market in every day products, with an attractive portfolio of properties for emerging applications ranging from health care and leisure to electronics, cosmetics, energy, agriculture, food and environmental. However, the novelty in properties and forms of aerogels makes the development of a legislative framework particularly challenging for ensuring the safe development and use of nano-enabled products. The presented safety regulatory Compendium intends to share knowledge with the international aerogels community, as well as end-users and stakeholders, on the regulatory and safe handling procedures, as best safety practices, to be followed during the production process, handling, transport and end-use of aerogel-based formulations to mitigate human and environmental risks considering lack of data availability for this purpose in general.

Immune Portrayal of a New Therapy Targeting Microbiota in an Animal Model of Psoriasis.

BACKGROUND: Despite all the available treatments, psoriasis remains incurable; therefore, finding personalized therapies is a continuous challenge. Psoriasis is linked to a gut microbiota imbalance, highlighting the importance of the gut-skin axis and its inflammatory mediators. Restoring this imbalance can open new perspectives in psoriasis therapy. We investigated the effect of purified IgY raised against pathological human bacteria antibiotic-resistant in induced murine psoriatic dermatitis (PSO). METHODS: To evaluate the immune portrayal in an imiquimod experimental model, before and after IgY treatment, xMAP array and flow cytometry were used. RESULTS: There were significant changes in IL-1α,ß, IL-5, IL-6, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17a, IFN-γ, TNF-α, IP-10/CXCL10, MCP-1/CCL2, MIP-1α/CCL3, MIP-1ß/CCL4, MIG/CXCL9, and KC/CXCL1 serum levels. T (CD3ε+), B (CD19+) and NK (NK1.1+) cells were also quantified. In our model, TNF-α, IL-6, and IL-1ß cytokines and CXCL1 chemokine have extremely high circulatory levels in the PSO group. Upon experimental therapy, the cytokine serum values were not different between IgY-treated groups and spontaneously remitted PSO. CONCLUSIONS: Using the murine model of psoriatic dermatitis, we show that the orally purified IgY treatment can lead to an improvement in skin lesion healing along with the normalization of cellular and humoral immune parameters.

The 3D Printing and Evaluation of Surgical Guides with an Incorporated Irrigation Channel for Dental Implant Placement.

Dental implant insertion requires the preparation of the implant bed via surgical drilling. During this stage, irrigation is essential to avoid thermal damage to the surrounding bone. Surgical guides enhance the accuracy of the implant site preparation, but they mask the drilling site, hampering coolant delivery. A variety of designs are aimed at improving the coolant access to the target site. Using standard dental implant simulation software, this paper presents an in-house design and 3D printing workflow for building surgical guides that incorporate a coolant channel directed toward the entry point of the burr. The proposed design was evaluated in terms of the bone temperature elevations caused by drilling performed at 1500 rpm, under an axial load of 2 kg, and irrigation with 40 mL/min of saline solution at 25 °C. Temperature measurements were performed on porcine femoral pieces, in the middle of the cortical bone layer, at 1 mm from the edge of the osteotomy. The mean temperature rise was 3.2 °C for a cylindrical sleeve guide, 2.7 °C for a C-shaped open-sleeve guide, and 2.1 °C for the guide with an incorporated coolant channel. According to a one-way ANOVA, the differences between these means were marginally insignificant (p = 0.056). The individual values of the peak temperature change remained below the bone damage threshold (10 °C) in all cases. Remarkably, the distribution of the recorded temperatures was the narrowest for the guide with internal irrigation, suggesting that, besides the most effective cooling, it provides the most precise control of the intraosseous temperature. Further studies could test different design variants, experimental models (including live animals), and might involve computer simulations of the bone temperature field.

Looking into the Skin in Health and Disease: From Microscopy Imaging Techniques to Molecular Analysis.

The skin is a complex organ that includes a wide variety of tissue types with different embryological origins [...].

Insights into Nutritional Strategies in Psoriasis.

Psoriasis, an autoimmune chronic inflammatory skin condition, has a high incidence in the general population, reaching 2-4%. Its pathogenesis involves an interplay of genetic factors, immune disturbances, and environmental factors. Within the environmental factors that aid the appearance of this autoimmune skin disease, the Western lifestyle and overall diet play important roles in the steady growth in psoriasis prevalence. Furthermore, psoriasis is associated with comorbidities such as psoriatic arthritis, cardiovascular disease, metabolic syndrome, and obesity. Accumulating evidence suggests that obesity is an important risk factor for psoriasis. Moreover, obesity aggravates established psoriasis, and a reduction in the body mass index can improve the clinical outcomes of psoriasis and increase the efficacy of standard psoriasis therapies. The possible connection between this autoimmune disease and obesity relies on the fact that white adipose tissue is an essential endocrine organ that secretes an array of immune mediators and inflammatory and metabolic factors with pro-inflammatory action. Thus, immune-mediated mechanisms in both psoriasis and obesity conditions are common factors. This paper describes the factors that link obesity with skin autoimmune disease and highlights the importance of the stimulatory or regulatory effects of nutrients and food in psoriasis and the possible improvement of psoriasis through nutritional strategies.

Learning Effects in Air Displacement Plethysmography.

Air displacement plethysmography (ADP) is a widespread technique for assessing global obesity in both health and disease. The reliability of ADP has been demonstrated by studies focused on duplicate trials. The present study was purported to evaluate learning effects on the reliability of body composition assessment using the BOD POD system, the sole commercially available ADP instrument. To this end, quadruplicate trials were performed on a group of 105 subjects (51 women and 54 men). We estimated measurement error from pairs of consecutive trials-(1,2), (2,3), and (3,4)-to test the hypothesis that early measurements are subject to larger errors. Indeed, statistical analysis revealed that measures of reliability inferred from the first two trials were inferior to those computed for the other pairs of contiguous trials: for percent body fat (%BF), the standard error of measurement (SEM) was 1.04% for pair (1,2), 0.71% for pair (2,3), and 0.66% for pair (3,4); the two-way random effects model intraclass correlation coefficient (ICC) was 0.991 for pair (1,2), and 0.996 for pairs (2,3) and (3,4). Our findings suggest that, at least for novice subjects, the first ADP test should be regarded as a practice trial. When the remaining trials were pooled together, the reliability indices of single ADP tests were the following: ICC = 0.996, SEM = 0.70%, and minimum detectable change (MDC) = 1.93% for %BF, and ICC = 0.999, SEM = 0.49 kg, and MDC = 1.35 kg for fat-free mass (FFM). Thus, the present study pleads for eliminating learning effects to further increase the reliability of ADP.

Hollow Particles Obtained by Prilling and Supercritical Drying as a Potential Conformable Dressing for Chronic Wounds.

The production of aerogels for different applications has been widely known, but the use of polysaccharide-based aerogels for pharmaceutical applications, specifically as drug carriers for wound healing, is being recently explored. The main focus of this work is the production and characterization of drug-loaded aerogel capsules through prilling in tandem with supercritical extraction. In particular, drug-loaded particles were produced by a recently developed inverse gelation method through prilling in a coaxial configuration. Particles were loaded with ketoprofen lysinate, which was used as a model drug. The core-shell particles manufactured by prilling were subjected to a supercritical drying process with CO2 that led to capsules formed by a wide hollow cavity and a tunable thin aerogel layer (40 µm) made of alginate, which presented good textural properties in terms of porosity (89.9% and 95.3%) and a surface area up to 417.0 m2/g. Such properties allowed the hollow aerogel particles to absorb a high amount of wound fluid moving very quickly (less than 30 s) into a conformable hydrogel in the wound cavity, prolonging drug release (till 72 h) due to the in situ formed hydrogel that acted as a barrier to drug diffusion.

Hyaluronan and Reactive Oxygen Species Signaling-Novel Cues from the Matrix?

Hyaluronan (HA) is a naturally occurring non-sulfated glycosaminoglycan (GAG) localized to the cell surface and the tissue extracellular matrix (ECM). It is composed of disaccharides containing glucuronic acid and N-acetylglucosamine, is synthesized by the HA synthase (HAS) enzymes and is degraded by hyaluronidase (HYAL) or reactive oxygen and nitrogen species (ROS/RNS) actions. HA is deposited as a high molecular weight (HMW) polymer and degraded to low molecular weight (LMW) fragments and oligosaccharides. HA affects biological functions by interacting with HA-binding proteins (hyaladherins). HMW HA is anti-inflammatory, immunosuppressive, and antiangiogenic, whereas LMW HA has pro-inflammatory, pro-angiogenetic, and oncogenic effects. ROS/RNS naturally degrade HMW HA, albeit at enhanced levels during tissue injury and inflammatory processes. Thus, the degradation of endothelial glycocalyx HA by increased ROS challenges vascular integrity and can initiate several disease progressions. Conversely, HA exerts a vital role in wound healing through ROS-mediated HA modifications, which affect the innate immune system. The normal turnover of HA protects against matrix rigidification. Insufficient turnover leads to increased tissue rigidity, leading to tissue dysfunction. Both endogenous and exogenous HMW HA have a scavenging capacity against ROS. The interactions of ROS/RNS with HA are more complex than presently perceived and present an important research topic.

Using A-Mode Ultrasound to Assess the Body Composition of Soccer Players: A Comparative Study of Prediction Formulas.

For elite athletes, monitoring body composition is important for maximizing performance without health risks. Amplitude (A)-mode ultrasound (AUS) has attracted increasing attention as an alternative to skinfold thickness measurements commonly used for assessing the amount of body fat in athletes. AUS accuracy and precision, however, depend on the formula used to predict body fat percentage (%BF) from subcutaneous fat layer thicknesses. Therefore, this study evaluates the accuracy of the 1-point biceps (B1), 9-sites Parrillo, 3-sites Jackson and Pollock (JP3), and 7-sites Jackson and Pollock (JP7) formulas. Relying on the previous validation of the JP3 formula in college-aged male athletes, we took AUS measurements in 54 professional soccer players (aged 22.9 ± 3.83 y, mean ± SD) and compared the results given by different formulas. The Kruskal-Wallis test indicated significant differences (p < 10-6), and Conover's post hoc test revealed that the JP3 and JP7 data come from the same distribution, whereas the data given by B1 and P9 differ from all the others. Lin's concordance correlation coefficients for B1 vs. JP7, P9 vs. JP7, and JP3 vs. JP7 were 0.464, 0.341, and 0.909, respectively. The Bland-Altman analysis indicated mean differences of -0.5 %BF between JP3 and JP7, 4.7 %BF between P9 and JP7, and 3.1 %BF between B1 and JP7. This study suggests that JP7 and JP3 are equally valid, whereas P9 and B1 overestimate %BF in athletes.

Skin Cancer Pathobiology at a Glance: A Focus on Imaging Techniques and Their Potential for Improved Diagnosis and Surveillance in Clinical Cohorts.

Early diagnosis is essential for completely eradicating skin cancer and maximizing patients' clinical benefits. Emerging optical imaging modalities such as reflectance confocal microscopy (RCM), optical coherence tomography (OCT), magnetic resonance imaging (MRI), near-infrared (NIR) bioimaging, positron emission tomography (PET), and their combinations provide non-invasive imaging data that may help in the early detection of cutaneous tumors and surgical planning. Hence, they seem appropriate for observing dynamic processes such as blood flow, immune cell activation, and tumor energy metabolism, which may be relevant for disease evolution. This review discusses the latest technological and methodological advances in imaging techniques that may be applied for skin cancer detection and monitoring. In the first instance, we will describe the principle and prospective clinical applications of the most commonly used imaging techniques, highlighting the challenges and opportunities of their implementation in the clinical setting. We will also highlight how imaging techniques may complement the molecular and histological approaches in sharpening the non-invasive skin characterization, laying the ground for more personalized approaches in skin cancer patients.

Mathematical and computational models in spheroid-based biofabrication.

Ubiquitous in embryonic development, tissue fusion is of interest to tissue engineers who use tissue spheroids or organoids as building blocks of three-dimensional (3D) multicellular constructs. This review presents mathematical models and computer simulations of the fusion of tissue spheroids. The motivation of this study stems from the need to predict the post-printing evolution of 3D bioprinted constructs. First, we provide a brief overview of differential adhesion, the main morphogenetic mechanism involved in post-printing structure formation. It will be shown that clusters of cohesive cells behave as an incompressible viscous fluid on the time scale of hours. The discussion turns then to mathematical models based on the continuum hydrodynamics of highly viscous liquids and on statistical mechanics. Next, we analyze the validity and practical use of computational models of multicellular self-assembly in live constructs created by tissue spheroid bioprinting. Finally, we discuss the perspectives of the field as machine learning starts to reshape experimental design, and modular robotic workstations tend to alleviate the burden of repetitive tasks in biofabrication. STATEMENT OF SIGNIFICANCE: Bioprinted constructs are living systems, which evolve via morphogenetic mechanisms known from developmental biology. This review presents mathematical and computational tools devised for modeling post-printing structure formation. They help achieving a desirable outcome without expensive optimization experiments. While previous reviews mainly focused on assumptions, technical details, strengths, and limitations of computational models of multicellular self-assembly, this article discusses their validity and practical use in biofabrication. It also presents an overview of mathematical models that proved to be useful in the evaluation of experimental data on tissue spheroid fusion, and in the calibration of computational models. Finally, the perspectives of the field are discussed in the advent of robotic biofabrication platforms and bioprinting process optimization by machine learning.

Bio-Based Aerogels for the Removal of Heavy Metal Ions and Oils from Water: Novel Solutions for Environmental Remediation.

Contamination of the aqueous environment caused by the presence of heavy metal ions and oils is a growing concern that must be addressed to reduce their detrimental impact on living organisms and safeguard the environment. Recent efficient and environmentally friendly remediation methods for the treatment of water are based on third-generation bioaerogels as emerging applications for the removal of heavy metal ions and oils from aqueous systems. The peculiarities of these materials are various, considering their high specific surface area and low density, together with a highly porous three-dimensional structure and tunable surface chemistry. This review illustrates the recent progress in aerogels developed from cellulose and chitosan as emerging materials in water treatment. The potential of aerogel-based adsorbents for wastewater treatment is reported in terms of adsorption efficacy and reusability. Despite various gaps affecting the manufacturing and production costs of aerogels that actually limit their successful implementation in the market, the research progress suggests that bio-based aerogels are ready to be used in water-treatment applications in the near future.

Langerhans Cells-Revising Their Role in Skin Pathologies.

Langerhans cells (LCs) constitute a cellular immune network across the epidermis. Because they are located at the skin barrier, they are considered immune sentinels of the skin. These antigen-presenting cells are capable of migrating to skin draining lymph nodes to prime adaptive immune cells, namely T- and B-lymphocytes, which will ultimately lead to a broad range of immune responses. Moreover, LCs have been shown to possess important roles in the anti-cancer immune responses. Indeed, the literature nicely highlights the role of LCs in melanoma. In line with this, LCs have been found in melanoma tissues where they contribute to the local immune response. Moreover, the immunogenic properties of LCs render them attractive targets for designing vaccines to treat melanoma and autoimmune diseases. Overall, future studies will help to enlarge the portfolio of immune properties of LCs, and aid the prognosis and development of novel therapeutic approaches to treating skin pathologies, including cancers.

Re-emerging concepts of immune dysregulation in autism spectrum disorders.

Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by communication and social interaction deficits, and by restricted interests and stereotyped, repetitive behavior patterns. ASD has a strong genetic component and a complex architecture characterized by the interplay of rare and common genetic variants. Recently, increasing evidence suggest a significant contribution of immune system dysregulation in ASD. The present paper reviews the latest updates regarding the altered immune landscape of this complex disorder highlighting areas with potential for biomarkers discovery as well as personalization of therapeutic approaches. Cross-talk between the central nervous system and immune system has long been envisaged and recent evidence brings insights into the pathways connecting the brain to the immune system. Disturbance of cytokine levels plays an important role in the establishment of a neuroinflammatory milieu in ASD. Several other immune molecules involved in antigen presentation and inflammatory cellular phenotypes are also at play in ASD. Maternal immune activation, the presence of brain-reactive antibodies and autoimmunity are other potential prenatal and postnatal contributors to ASD pathophysiology. The molecular players involved in oxidative-stress response and mitochondrial system function, are discussed as contributors to the pro-inflammatory pattern. The gastrointestinal inflammation pathways proposed to play a role in ASD are also discussed. Moreover, the body of evidence regarding some of the genetic factors linked to the immune system dysregulation is reviewed and discussed. Last, but not least, the epigenetic traits and their interactions with the immune system are reviewed as an expanding field in ASD research. Understanding the immune-mediated pathways that influence brain development and function, metabolism, and intestinal homeostasis, may lead to the identification of robust diagnostic or predictive biomarkers for ASD individuals. Thus, novel therapeutic approaches could be developed, ultimately aiming to improve their quality of life.

Healthy Ageing Reflected in Innate and Adaptive Immune Parameters.

Purpose: The aim of the paper is to establish and quantify the relation between healthy ageing and the innate and adaptive immune parameters as indicators of age-related diseases. Patients: In order to observe the immunological changes that occur according to age, several humoral and cellular immune parameters were investigated for 288 healthy donors (30-80 years). Subjects' selection was done using clinical, biochemical and immunological parameters of inclusion/exclusion criteria from SENIEUR protocol. Results: Age-related changes were observed for both humoral and cellular immune parameters. Lymphocyte immunophenotyping revealed several significant differences in the distribution of cells, both intra- and inter-age groups, namely decreased values of T-CD3+, T-CD8+ and NK cells, and elevated values for T-CD4+, T-CD4+/T-CD8+ ratio and B cells. The percentages of unstimulated neutrophils that show basal oxidative activity and the intensity of this activity had an increasing tendency age-related. The percentage of N-Formyl-Methionyl-Leucyl-Phenylalanine stimulated neutrophils clearly decreases with age, and is associated with an increasing intensity of oxidative activity. Our data also have shown an increased percentage of oxidative neutrophils after phorbol 12-myristate 13-acetate stimulation and an elevated oxidative activity with age. Conclusion: Overall healthy ageing is governed by some immune-related deregulations that account for immune exhaustion due to numerous developed immune processes during a life-time and the age-related diseases.

Skin Inflammation-A Cornerstone in Dermatological Conditions.

The skin provides more than a simple mechanical barrier against external aggressors; it is an array of effector cells and molecules that constitute the skin's immune system, as defined by Bos and Kapsenberg in 1986 [...].