Additional file 1: Figure S1. of Control of Alginate Core Size in Alginate-Poly (Lactic-Co-Glycolic) Acid Microparticles

The standard curve of fluorescence vs calcein concentration. Fluorescence with emission/excitation of 485/535nm = {3.82 × [calcein concentration, µM]2} + {67.16 × [calcein concentration, µM]} was obtained with coefficient of determination, R2 = 0.997

Additional file 3: Figure S3. of Control of Alginate Core Size in Alginate-Poly (Lactic-Co-Glycolic) Acid Microparticles

Calcein encapsulation efficiency for PLGA microparticles containing different PLGA core sizes. PLGA microparticles demonstrated lower encapsulation efficiency as compared to alginate-PLGA microparticles in Additional file 2

Additional file 2: Figure S2. of Control of Alginate Core Size in Alginate-Poly (Lactic-Co-Glycolic) Acid Microparticles

Calcein encapsulation efficiency for alginate-PLGA microparticles containing different sizes of alginate core. Alginate-PLGA microparticles containing 10 and 50Âľm alginate cores (homo-mag and vort-mag) demonstrated the high calcein encapsulation efficiency as compared to mag-mag alginate-PLGA microparticles.

MOESM1 of Microneedle physical contact as a therapeutic for abnormal scars

Additional file 1: Figure S1. Microneedle contact and material cytotoxicity on fibroblasts. A) Representative phase contrast, blue and red fluorescence images of control and microneedle-treated normal fibroblasts (NF) incubated with Hoechst 33342 (blue) and propidium iodide (red). B) Fraction (%) of dead cells to total cells for the following studied groups: untreated control of normal fibroblasts (NF_C), normal fibroblasts cultured in presence of the microneedles but avoiding direct contact with the needles (NF_LCP) and microneedle-treated normal fibroblasts (NF_microneedle). The group NF_LCP aimed to evaluate the intrinsic toxicity of the material. For this, the microneedles were fixated to the walls of the well of the cell culture plate and immersed on the medium (schematically represented in B). C) Comparison of untreated control and microneedle-treated cells for keloids fibroblasts (KF) and normal fibroblasts (NF). Statistical significance has been indicated with *p < 0.01. Figure S2. Microneedles induce increased cell death. A) Representative images and B) quantification of dead Keloid Fibroblast (KF) cells per total cells in untreated controls and microneedle-treated cells. Quantification (B) was performed within the indicated (dotted lines) region of interest (ROI). The ROI was selected based on the region in which the microneedles were applied. This ROI from microneedle-treated samples was overlaid on the images of the untreated controls in order to have comparable ROI for quantification purposes. The used microscopic images were taken under identical fluorescence conditions and using the same magnification. Dead cells are propidium (PI—red)-labeled and total cells are Hoechst 33342 (blue)-labeled. Statistical significances are indicated with **p < 0.01, N = 3, values are mean ± SD. Figure S3. Quantified microneedle efficacy through the scar elevation index. A) SEI—scar elevation index measured on untreated & microneedle treated rabbit ear wounds. Representative untreated (B) and microneedle treated (C) wounds with the (---) region demarcating the raised neodermis and the (―) region signifying the original boundary where the wound was inflicted. *P < 0.01, N = 3

Table_1_Immune-mediated diseases are associated with a higher risk of ALS incidence: a prospective cohort study from the UK Biobank.docx

ObjectiveThe occurrence of immune-mediated diseases (IMDs) in amyotrophic lateral sclerosis (ALS) patients is widely reported. However, whether IMDs and ALS is a simple coexistence or if there exists causal relationships between the two has been a subject of great interest to researchers.MethodsA total of 454,444 participants from the prospective cohort of UK Biobank were recruited to investigate the longitudinal association between IMDs and ALS. Previously any IMDs and organ specific IMDs were analyzed in relation to the following incident ALS by Cox-proportional hazard models. Subgroup analyses were performed to explore the covariates of these relationships.ResultsAfter adjusting for potential covariates, the multivariate analysis showed that any IMDs were associated with an increased risk of ALS incidence (HR:1.42, 95%CI:1.03-1.94). IMDs of the endocrine-system and the intestinal-system were associated with increased risk of ALS incidence (endocrine-system IMDs: HR:3.01, 95%CI:1.49-6.06; intestinal system IMDs: HR:2.07, 95%CI: 1.14-3.77). Subgroup analyses revealed that immune burden, including IMD duration and the severity of inflammation had specific effects on the IMD-ALS association. In participants with IMD duration≥10 years or CRP≥1.3mg/L or females, previous IMDs increased the risk of incident ALS; however, in participants with IMD duration InterpretationOur study provides evidence that previous any IMDs and endocrine-system and the intestinal-system specific IMDs are associated with an increased risk of developing ALS in females, but not in males.</p

Smart Magnetic Nanosensors Synthesized through Layer-by-Layer Deposition of Molecular Beacons for Noninvasive and Longitudinal Monitoring of Cellular mRNA

Noninvasive and longitudinal monitoring of gene expression in living cells is essential for understanding and monitoring cellular activities. Herein, a smart magnetic nanosensor is constructed for the real-time, noninvasive, and longitudinal monitoring of cellular mRNA expression through the layer-by-layer deposition of molecular beacons (MBs) and polyethylenimine on the iron oxide nanoparticles. The loading of MBs, responsible for the signal intensity and the tracking time, was easily tuned with the number of layers incorporated. The idea was first demonstrated with the magnetic nanosensors for glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA, which was efficiently internalized into the cells under the influence of magnetic field. This nanosensor allowed the continuous monitoring of the cellular GAPDH mRNA expression for 1 month. Then this platform was further utilized to incorporate two kinds of MBs for alkaline phosphatase (ALP) and GAPDH mRNAs, respectively. The multifunctional nanosensors permitted the simultaneous monitoring of the reference gene (GAPDH mRNA) and the early osteogenic differentiation marker (ALP mRNA) expression. When the fluorescence signal ratio between ALP mRNA MBs and GAPDH mRNA MBs was taken, the dynamic osteogenic differentiation process of MSCs was accurately monitored

Nitric Oxide Nanosensors for Predicting the Development of Osteoarthritis in Rat Model

Osteoarthritis (OA) is a chronic arthritic disease that causes the overproduction of inflammatory factors such as nitric oxide (NO). This study develops a NO nanosensor to predict the OA development. The nanosensor is synthesized by encapsulating the NO sensing molecules (i.e., 4-amino-5-methylamino-2′,7′-difluorofluorescein Diaminofluorescein-FM (DAF-FM)) within the biodegradable poly­(lactic-<i>co</i>-glycolic acid) nanoparticles. In vitro, the nanosensor allows the monitoring of the NO release in interleukin-1β-stimulated chondrocytes and the alleviated effect of N^G-monomethyl-l-arginine (a NO inhibitor) and andrographolide (an anti-inflammatory agent). In the rat OA model, it permits the quantification of NO level in joint fluid. The proposed NO nanosensor may facilitate a noninvasive and real-time evaluation of the OA development

Iron Oxide Nanoparticle-Powered Micro-Optical Coherence Tomography for in Situ Imaging the Penetration and Swelling of Polymeric Microneedles in the Skin

In recent years, polymeric microneedles (MNs) have attracted keen interests among researchers because of their applicability in transdermal drug delivery and interstitial skin fluid (ISF) extraction. When designing and characterizing such devices, it is critical to monitor their real-time in vitro and in vivo performances to optimize the desired effects, yet most of the existing methods are incapable of such functions. To address this unmet need, we develop a real-time noninvasive imaging methodology by integrating iron oxide (Fe₃O₄) nanoparticles into polymeric MNs to enhance image contrast for micro-optical coherence tomography (μOCT) imaging. Using the Fe₃O₄-integrated polystyrene-<i>block</i>-poly­(acrylic acid) (PS-<i>b</i>-PAA) MNs as an example, we evaluate the influences of Fe₃O₄ concentrations on contrast enhancement in μOCT imaging and visualize the real-time swelling process of polymeric MNs in biological samples for the first time. Our results show that a concentration of ∼4–5 wt % Fe₃O₄ nanoparticles not only helps achieve the best contrast-to-noise ratio in μOCT imaging, which is 10 times higher than that without Fe₃O₄ nanoparticles in air and hydrogel, but also enables the real-time changes in the profile of MNs to be observed clearly in their swelling process in skin tissues. On the basis of such findings, we utilize the optimized concentration of Fe₃O₄ nanoparticles to further quantitatively study the swelling kinetics of PS-<i>b</i>-PAA MNs in agarose hydrogel and fresh skin tissues, which lasts ∼20 and ∼30–35 s, respectively. The suitability of such a methodology for enhancing μOCT imaging would greatly facilitate the development and clinical translation of MN-based medical technologies

Interference-free Micro/nanoparticle Cell Engineering by Use of High-Throughput Microfluidic Separation

Engineering cells with active-ingredient-loaded micro/nanoparticles is becoming increasingly popular for imaging and therapeutic applications. A critical yet inadequately addressed issue during its implementation concerns the significant number of particles that remain unbound following the engineering process, which inadvertently generate signals and impart transformative effects onto neighboring nontarget cells. Here we demonstrate that those unbound micro/nanoparticles remaining in solution can be efficiently separated from the particle-labeled cells by implementing a fast, continuous, and high-throughput Dean flow fractionation (DFF) microfluidic device. As proof-of-concept, we applied the DFF microfluidic device for buffer exchange to sort labeled suspension cells (THP-1) from unbound fluorescent dye and dye-loaded micro/nanoparticles. Compared to conventional centrifugation, the depletion efficiency of free dyes or particles was improved 20-fold and the mislabeling of nontarget bystander cells by free particles was minimized. The microfluidic device was adapted to further accommodate heterogeneous-sized mesenchymal stem cells (MSCs). Complete removal of unbound nanoparticles using DFF led to the usage of engineered MSCs without exerting off-target transformative effects on the functional properties of neighboring endothelial cells. Apart from its effectiveness in removing free particles, this strategy is also efficient and scalable. It could continuously process cell solutions with concentrations up to 10⁷ cells·mL^–1 (cell densities commonly encountered during cell therapy) without observable loss of performance. Successful implementation of this technology is expected to pave the way for interference-free clinical application of micro/nanoparticle engineered cells

Supplemental material for Detection of Bacteria in Water with β-Galactosidase-Coated Magnetic Nanoparticles

<p>Supplemental material for Detection of Bacteria in Water with β-Galactosidase-Coated Magnetic Nanoparticles by Mingyue Cui, Hao Chang, Yang Zhong, Min Wang, Tianze Wu, Xiao Hu, Zhichuan J. Xu, and Chenjie Xu in SLAS Technology</p