Oscillations of a soft viscoelastic drop

A soft viscoelastic drop has dynamics governed by the balance between surface tension, viscosity, and elasticity, with the material rheology often being frequency dependent, which are utilized in bioprinting technologies for tissue engineering and drop-deposition processes for splash suppression. We study the free and forced oscillations of a soft viscoelastic drop deriving (1) the dispersion relationship for free oscillations, and (2) the frequency response for forced oscillations, of a soft material with arbitrary rheology. We then restrict our analysis to the classical cases of a Kelvin–Voigt and Maxwell model, which are relevant to soft gels and polymer fluids, respectively. We compute the complex frequencies, which are characterized by an oscillation frequency and decay rate, as they depend upon the dimensionless elastocapillary and Deborah numbers and map the boundary between regions of underdamped and overdamped motions. We conclude by illustrating how our theoretical predictions for the frequency-response diagram could be used in conjunction with drop-oscillation experiments as a “drop vibration rheometer”, suggesting future experiments using either ultrasonic levitation or a microgravity environment.

Effect of Substrate Stiffness on Human Intestinal Enteroids Infectivity by Enteroaggregative Escherichia coli

Human intestinal enteroids (HIE) models have contributed significantly to our understanding of diarrheal diseases and other intestinal infections, but their routine culture conditions fail to mimic the mechanical environment of the native intestinal wall. Because the mechanical characteristics of the intestine significantly alter how pathogens interact with the intestinal epithelium, we used different concentrations of polyethylene glycol (PEG) to generate soft (~2 kPa), medium (~10 kPa), and stiff (~100 kPa) hydrogel biomaterial scaffolds. The height of HIEs cultured in monolayers atop these hydrogels was 18 μm whereas HIEs grown on rigid tissue culture surfaces (with stiffness in the GPa range) were 10 μm. Substrate stiffness also influenced the amount of enteroaggregative E. coli (EAEC strain 042) adhered to the HIEs. We quantified a striking difference in adherence pattern; on the medium and soft gels, the bacteria formed clusters of >100 and even >1000 on both duodenal and jejunal HIEs (such as would be found in biofilms), but did not on glass slides and stiff hydrogels. All hydrogel cultured HIEs showed significant enrichment for gene and signaling pathways related to epithelial differentiation, cell junctions and adhesions, extracellular matrix, mucins, and cell signaling compared to the HIEs cultured on rigid tissue culture surfaces. Collectively, these results indicate that the HIE monolayers cultured on the hydrogels are primed for a robust engagement with their mechanical environment, and that the soft hydrogels promote the formation of larger EAEC aggregates, likely through an indirect differential effect on mucus.

Efficient Matrix Cleanup of Soft-Gel-Type Dietary Supplements for Rapid Screening of 92 Illegal Adulterants Using EMR-Lipid dSPE and UHPLC-Q/TOF-MS

An efficient matrix cleanup method was developed for the rapid screening of 92 illegal adulterants (25 erectile dysfunction drugs, 15 steroids, seven anabolic steroids, 12 antihistamines, 12 nonsteroidal anti-inflammatory drugs (NSAIDs), four diuretics, and 17 weight-loss drugs) in soft-gel-type supplements by ultra-high performance liquid chromatography-quadrupole/time of flight-mass spectrometry (UHPLC-Q/TOF-MS). As representative green chemistry methods, three sample preparation methods (dispersive liquid-liquid microextraction (DLLME), “quick, easy, cheap, effective, rugged, and safe” dispersive solid-phase extraction (QuEChERS-dSPE), and enhanced matrix removal-lipid (EMR-Lipid) dSPE) were evaluated for matrix removal efficiency, recovery rate, and matrix effect. In this study, EMR-Lipid dSPE was shown to effectively remove complicated matrix contents in soft-gels, compared to DLLME and QuEChERS-dSPE. For the rapid screening of a wide range of adulterants, extracted common ion chromatogram (ECIC) and neutral loss scan (NLS) based on specific common MS/MS fragments were applied to randomly collected soft-gel-type dietary supplement samples using UHPLC-Q/TOF-MS. Both ECICs and NLSs enabled rapid and simple screening of multi-class adulterants and could be an alternative to the multiple reaction monitoring (MRM) method. The developed method was validated in terms of limit of detection (LOD), precision, accuracy, recovery, and matrix effects. The range of LODs was 0.1–16 ng/g. The overall precision values were within 0.09–14.65%. The accuracy ranged from 81.6% to 116.6%. The recoveries and matrix effects of 92 illegal adulterants ranged within 16.9–119.4% and 69.8–114.8%, respectively. The established method was successfully applied to screen and identify 92 illegal adulterants in soft-gels. This method can be a promising tool for the high-throughput screening of various adulterants in dietary supplements and could be used as a more environmentally friendly routine analytical method for screening dietary supplements illegally adulterated with multi-class drug substances.

Comparison of Label Claims, Measured Curcuminoid Content and Dosage Form Performance Quality With Industry Standards for Turmeric Dietary Supplements

Objectives Turmeric (Curcuma longa) is a popular ingredient in dietary supplements (DS), promoted for a variety of health effects. FDA label regulations require information on the total weight of each botanical or extract present in a botanical DS, unless it is part of a proprietary blend. Information on the concentration of phytochemicals in extracts is voluntary. DS with turmeric rhizome powders and/or extracts were tested for curcuminoid content and the results compared to label claims and industry standards as part of the Dietary Supplement Ingredient Database (DSID) research program. The performance quality of the dosage forms was also evaluated. Methods Commonly consumed DS (n = 54 × 2 lots) with turmeric as the only or primary ingredient were analyzed for curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Certified reference materials, in-house controls and blind sample duplicates were measured to ensure the quality of results from three laboratories. Percentages of total curcuminoids (TC) and ratios of the three curcuminoids were compared to published standards. Voluntary content claims were categorized as “complete” (all listed turmeric ingredients had a TC claim) or “partial” (only one of two had a TC claim). Tablets/caplets (n = 9) and capsules (n = 39) were also tested for disintegration and soft gels (n = 4) for rupture. Results Measured TC amounts varied widely among products. At the most common labeled level of 500 mg of turmeric per day, measured TC results ranged from 16 to 554 mg (n = 12). DS with voluntary claims for TC (n = 41) had analytical content averaging 6.2% above label, and 63% within ± 10% of claims. DS with complete TC claims had significantly higher levels in mg per day than those with partial or no claims (735 ± 87, 137 ± 37, 196 ± 57; mean ± SE, respectively). The curcuminoid ratios and/or the percentages of TC differed significantly from United States Pharmacopeial (USP) standards for at least 10 DS. Most (87.5%) DS passed disintegration tests and all soft gels passed the rupture test. Conclusions Current labeling requirements for turmeric DS may be insufficient to inform researchers and consumers about the actual content of TC in DS because the extract concentrations vary widely. Voluntary claims for TC amounts were reasonably accurate. Most turmeric DS met USP disintegration and composition standards. Funding Sources NIH-ODS and USDA-ARS.

Engineering cartilage graft using mesenchymal stem cell laden polyacrylamide-galactoxyloglucan hydrogel for transplantation

Hydrogels are reported to have various biomedical field applications, and many reports also suggest that soft gels promote stem cell differentiation. Chondrogenic differentiation of mesenchymal stem cells (MSC) is significant in articular cartilage repair. This study focuses on polysaccharide-based hydrogels which enhance chondrocyte lineage differentiation of MSC when grown in the hydrogels. This study implies that the prepared hydrogels promote specific lineage without any external chemical induction factors. The techniques, including immunofluorescence and functional assays to assess the differentiation and in vivo implantation, were employed. All observations paved the way towards confirmation that the galactoxyloglucan-based hydrogel is an attractive candidate for supporting stem cell growth and cartilaginous differentiation.

Optogenetic control of apical constriction induces synthetic morphogenesis in mammalian tissues

During embryonic development, cellular forces synchronize in space and time to generate functional tissue shapes. Apical constriction is one of these force-generating processes, and it is necessary to modulate epithelial curvature in fundamental morphogenetic events, such as neural tube folding. The emerging field of synthetic developmental biology proposes bottom-up approaches to examine the contribution of each cellular process to complex morphogenesis. However, the shortage of tools to manipulate three-dimensional (3D) shapes of mammalian tissues currently hinders the progress of the field. Here we report the development of 'OptoShroom3', a new optogenetic tool that achieves fast spatiotemporal control of apical constriction in mammalian epithelia. Activation of OptoShroom3 through illumination of individual cells in an epithelial cell sheet reduced their apical surface while illumination of groups of cells caused deformation in the adjacent regions. By using OptoShroom3, we further manipulated 3D tissue shapes. Light-induced apical constriction provoked the folding of epithelial cell colonies on soft gels. Its application to murine and human neural organoids led to thickening of neuroepithelia, apical lumen reduction in optic vesicles, and flattening in neuroectodermal tissues. These results show that spatiotemporal control of apical constriction can trigger several types of 3D deformation depending on the initial tissue context.