Spatiotemporal monitoring of a periodontal multispecies biofilm model: demonstration of prebiotic treatment responses.

Biofilms are complex polymicrobial communities which are often associated with human infections such as the oral disease periodontitis. Studying these complex communities under controlled conditions requires in vitro biofilm model systems that mimic the natural environment as close as possible. This study established a multispecies periodontal model in the drip flow biofilm reactor in order to mimic the continuous flow of nutrients at the air-liquid interface in the oral cavity. The design is engineered to enable real-time characterization. A community of five bacteria, Streptococcus gordonii-GFPmut3*, Streptococcus oralis-GFPmut3*, Streptococcus sanguinis-pVMCherry, Fusobacterium nucleatum, and Porphyromonas gingivalis-SNAP26 is visualized using two distinct fluorescent proteins and the SNAP-tag. The biofilm in the reactor develops into a heterogeneous, spatially uniform, dense, and metabolically active biofilm with relative cell abundances similar to those in a healthy individual. Metabolic activity, structural features, and bacterial composition of the biofilm remain stable from 3 to 6 days. As a proof of concept for our periodontal model, the 3 days developed biofilm is exposed to a prebiotic treatment with L-arginine. Multifaceted effects of L-arginine on the oral biofilm were validated by this model setup. L-arginine showed to inhibit growth and incorporation of the pathogenic species and to reduce biofilm thickness and volume. Additionally, L-arginine is metabolized by Streptococcus gordonii-GFPmut3* and Streptococcus sanguinis-pVMCherry, producing high levels of ornithine and ammonium in the biofilm. In conclusion, our drip flow reactor setup is promising in studying spatiotemporal behavior of a multispecies periodontal community.ImportancePeriodontitis is a multifactorial chronic inflammatory disease in the oral cavity associated with the accumulation of microorganisms in a biofilm. Not the presence of the biofilm as such, but changes in the microbiota (i.e., dysbiosis) drive the development of periodontitis, resulting in the destruction of tooth-supporting tissues. In this respect, novel treatment approaches focus on maintaining the health-associated homeostasis of the resident oral microbiota. To get insight in dynamic biofilm responses, our research presents the establishment of a periodontal biofilm model including Streptococcus gordonii, Streptococcus oralis, Streptococcus sanguinis, Fusobacterium nucleatum, and Porphyromonas gingivalis. The added value of the model setup is the combination of simulating continuously changing natural mouth conditions with spatiotemporal biofilm profiling using non-destructive characterization tools. These applications are limited for periodontal biofilm research and would contribute in understanding treatment mechanisms, short- or long-term exposure effects, the adaptation potential of the biofilm and thus treatment strategies.

Role of Nanoparticles in Nanofluid Droplet Impact on Solid Surfaces.

Splashing of a liquid droplet onto a substrate, while ubiquitous, sits at the intersection of several key fluid mechanical regions. Typically, this problem is often simplified to the transition between spreading and splashing, even for splashing on complex surfaces. Recently, there has been increased interest in using not just pure liquids but also nanofluids in applications such as spray cooling. While the addition of a few percent of nanoparticles to a Newtonian fluid does not change its apparent viscosity, the influence of the nanoparticles on the splashing transition is pronounced. We often view splashing in terms of fluid mechanics where a simple material is subjected to a complex flow and the fluid can be simply characterized by a Newtonian viscosity. For nanofluids, we have an apparently simple material in a complex flow, but the results show that the impact of the particles is nontrivial. This implies that we must now combine some of the insights we obtain from studying the rheological properties of nanosuspensions with this already complex problem.

On the gelation of humins: from transient to covalent networks.

Humins are a by-product of many acid-catalyzed biorefinery processes converting polysaccharides into platform chemicals. The valorization of humin residue to increase the profit of biorefinery operations and reduce waste is a field that is growing interest as the production of humins continues to increase. This includes their valorization in materials science. For successful processing of humin-based materials, this study aims to understand the thermal polymerization mechanisms of humins from a rheological perspective. Thermal crosslinking of raw humins leads to an increase in their molecular weight, which in turn leads to the formation of a gel. Humin's gels structure combines physical (thermally reversible) and chemical (thermally irreversible) crosslinks, and temperature plays an essential role in the crosslink density and the gel properties. High temperatures delay the formation of a gel due to the scission of physicochemical interactions, drastically decreasing their viscosity, whereas upon cooling a stronger gel is formed combining the recovered physicochemical bonds and the newly created chemical crosslinks. Thus, a transition from a supramolecular network to a covalently crosslinked network is observed, and properties such as the elasticity or reprocessability of humin gels are influenced by the stage of polymerization.

Connecting particle clustering and rheology in attractive particle networks.

The structural properties of suspensions and other multiphase systems are vital to overall processability, functionality and acceptance among consumers. Therefore, it is crucial to understand the intrinsic connection between the microstructure of a material and the resulting rheological properties. Here, we demonstrate how the transitions in the microstructural conformations can be quantified and correlated to rheological measurements. We find semi-local parameters from graph theory, the mathematical study of networks, to be useful in linking structure and rheology. Our results, using capillary suspensions as a model system, show that the use of the clustering coefficient, in combination with the coordination number, is able to capture not only the agglomeration of particles, but also measures the formation of groups. These phenomena are tightly connected to the rheological properties. The present sparse networks cannot be described by established techniques such as betweenness centrality.

Negative normal stress differences N1-N2 in a low concentration capillary suspension.

Here, negative normal stress differences are reported in capillary suspensions, i.e. particle suspensions in a two-fluid system that creates strong capillary attractions, at a solid concentration of 25%, and a volume fraction that has heretofore been considered too low to show such normal stress differences. Such capillary suspensions have strong particle networks and are shear thinning for the entire range of shear rates studied. Capillary suspensions exist in two states: a pendular state when the secondary fluid preferentially wets the particles, and a capillary state when the bulk fluid is preferentially wetting. In the pendular state, the system undergoes a transition from a positive normal stress difference at high shear rates to a negative stress difference at low shear rates. These results are an indication of flexible flocs in the pendular state that are able to rotate to reorientate in the vorticity direction under shear. Analogous experiments were also conducted for the capillary state, where only a negative normal stress difference occurs. The capillary state system forms more network contacts due to droplet breakup at higher shear rates, which enhances the importance of hydrodynamic interactions in the non-colloidal suspension.

Fractal approaches to characterize the structure of capillary suspensions using rheology and confocal microscopy.

The rheological properties of a particle suspension can be substantially altered by adding a small amount of a secondary fluid that is immiscible with the bulk phase. The drastic change in the strength of these capillary suspensions arises due to the capillary forces, induced by the added liquid, leading to a percolating particle network. Using rheological scaling models, fractal dimensions are deduced from the yield stress and from oscillatory strain amplitude sweep data as function of the solid volume fraction. Exponents obtained using aluminum-oxide-based capillary suspensions, with a preferentially wetting secondary fluid, indicate an increase in the particle gel's fractal dimension with increasing particle size. This may be explained by a corresponding relative reduction in the capillary force compared to other forces. Confocal images using a glass model system show the microstructure to consist of compact particle flocs interconnected by a sparse backbone. Thus, using the rheological models two different fractal dimensionalities are distinguished - a lower network backbone dimension (D = 1.86-2.05) and an intrafloc dimension (D = 2.57-2.74). The latter is higher due to the higher local solid volume fraction inside of the flocs compared to the sparse backbone. Both of these dimensions are compared with values obtained by analysis of spatial particle positions from 3D confocal microscopy images, where dimensions between 2.43 and 2.63 are computed, lying between the two dimension ranges obtained from rheology. The fractal dimensions determined via this method corroborate the increase in structural compactness with increasing particle size.

Influence of mixing conditions on the rheological properties and structure of capillary suspensions.

The rheological properties of a suspension can be dramatically altered by adding a small amount of a secondary fluid that is immiscible with the bulk liquid. These capillary suspensions exist either in the pendular state where the secondary fluid preferentially wets the particles or the capillary state where the bulk fluid is preferentially wetting. The yield stress, as well as storage and loss moduli, depends on the size and distribution of secondary phase droplets created during sample preparation. Enhanced droplet breakup leads to stronger sample structures. In capillary state systems, this can be achieved by increasing the mixing speed and time of turbulent mixing using a dissolver stirrer. In the pendular state, increased mixing speed also leads to better droplet breakup, but spherical agglomeration is favored at longer times decreasing the yield stress. Additional mixing with a ball mill is shown to be beneficial to sample strength. The influence of viscosity variance between the bulk and second fluid on the droplet breakup is excluded by performing experiments with viscosity-matched fluids. These experiments show that the capillary state competes with the formation of Pickering emulsion droplets and is often more difficult to achieve than the pendular state.

Structure of Particle Networks in Capillary Suspensions with Wetting and Nonwetting Fluids.

The mechanical properties of a suspension can be dramatically altered by adding a small amount of a secondary fluid that is immiscible with the bulk phase. The substantial changes in the strength of these capillary suspensions arise due to the capillary force inducing a percolating particle network. Spatial information on the structure of the particle networks is obtained using confocal microscopy. It is possible, for the first time, to visualize the different types of percolating structures of capillary suspensions in situ. These capillary networks are unique from other types of particulate networks due to the nature of the capillary attraction. We investigate the influence of the three-phase contact angle on the structure of an oil-based capillary suspension with silica microspheres. Contact angles smaller than 90° lead to pendular networks of particles connected with single capillary bridges or clusters comparable to the funicular state in wet granular matter, whereas a different clustered structure, the capillary state, forms for angles larger than 90°. Particle pair distribution functions are obtained by image analysis, which demonstrate differences in the network microstructures. When porous particles are used, the pendular conformation also appears for apparent contact angles larger than 90°. The complex shear modulus can be correlated to these microstructural changes. When the percolating structure is formed, the complex shear modulus increases by nearly three decades. Pendular bridges lead to stronger networks than the capillary state network conformations, but the capillary state clusters are nevertheless much stronger than pure suspensions without the added liquid.

Capillary suspensions: Particle networks formed through the capillary force.

The addition of small amounts of a secondary fluid to a suspension can, through the attractive capillary force, lead to particle bridging and network formation. The capillary bridging phenomenon can be used to stabilize particle suspensions and precisely tune their rheological properties. This effect can even occur when the secondary fluid wets the particles less well than the bulk fluid. These materials, so-called capillary suspensions, have been the subject of recent research studying the mechanism for network formation, the properties of these suspensions, and how the material properties can be modified. Recent work in colloidal clusters is summarized and the relationship to capillary suspensions is discussed. Capillary suspensions can also be used as a pathway for new material design and some of these applications are highlighted. Results obtained to date are summarized and central questions that remain to be answered are proposed in this review.

Sex-specific substance abuse treatment for female healthcare professionals: implications.

OBJECTIVES: Gender plays a significant role in the development and treatment of substance abuse disorders. Sex-specific treatment for girls and women has recurrently proven more effective, with better outcomes than traditional treatment. Research on impaired healthcare professionals (HCPs) has largely focused on men, garnering little attention for women and sex differences. With the increasing numbers of female HCPs, it is imperative to identify potential sex differences that may have implications for treatment. Our study compared a convenience sample of male and female HCPs with substance abuse disorders treated in an outpatient program to identify sex differences that may have implications for treatment. METHODS: Our sample consisted of 96 HCPs (54 men, 42 women) and 17 non-healthcare professional (N-HCP) women. All of the participants were evaluated using the program's clinical interview and the Personality Assessment Inventory (PAI). Chart review data contained categorical variables, qualitative variables, diagnoses, and psychological test scores. A second analysis was conducted through two separate comparisons: the PAI results of comparing impaired female HCPs with impaired male HCPs and the PAI results of comparing impaired female HCPs with impaired female N-HCPs. RESULTS: Statistically significant differences indicated more male participants received prior treatment and more intensive treatment than female participants. More female subjects reported being diagnosed as having a comorbid psychiatric condition and taking psychotropic medications. Several statistically significant differences in the PAI scores were found. Among female HCPs, elevations were found in anxiety, depression, paranoia, and borderline personality disorder. Substantive differences, although not statistically significant, were elevations in somatic complaints and anxiety disorders in female HCPs. In the comparison of female HCPs and N-HCPs, the only statistically significant difference was the significantly higher anxiety score of N-HCPs. CONCLUSIONS: The results indicate greater differences between female HCPs and male HCPs than between female HCPs and N-HCPs.

Restructuring and aging in a capillary suspension.

The rheological properties of capillary suspensions, suspensions with small amounts of an added immiscible fluid, are dramatically altered with the addition of the secondary fluid. We investigate a capillary suspension to determine how the network ages and restructures at rest and under applied external shear deformation. The present work uses calcium carbonate suspended in silicone oil (11 % solids) with added water as a model system. Aging of capillary suspensions and their response to applied oscillatory shear is distinctly different from particulate gels dominated by the van der Waals forces. The suspensions dominated by the capillary force are very sensitive to oscillatory flow, with the linear viscoelastic regime ending at a deformation of only 0.1% and demonstrating power-law aging behavior. This aging persists for long times at low deformations or for shorter times with a sudden decrease in the strength at higher deformations. This aging behavior suggests that the network is able to rearrange and even rupture. This same sensitivity is not demonstrated in shear flow where very high shear rates are required to rupture the agglomerates returning the apparent viscosity of capillary suspensions to the same viscosity as for the pure vdW suspension. A transitional region is also present at intermediate water contents wherein the material response depends very strongly on the type, strength, and duration of the external forcing.

Enhanced contact flexibility from nanoparticles in capillary suspensions.

HYPOTHESIS: Sample-spanning particle networks are used to induce structure and a yield stress, necessary for 3D printing of porous ceramics and paints. In capillary suspensions, a small quantity of immiscible secondary fluid is incorporated into a suspension. By further adding nanoparticles with a range of hydrophobicities, the structure of the bridges and microparticle-microparticle contacts is expected to be modified, resulting in a tunable yield stress and shear moduli. Moreover, the compressibility of these samples, important in many processing and application steps, is expected to be sensitive to these changes. EXPERIMENT: The nanoparticle hydrophobicity was altered and their position relative to the microparticles and the bridges was examined using confocal microscopy where the correlation between bridge size and network structure was observed. A step-wise uniaxial compression test on the confocal was conducted to monitor the microparticle movement and structural changes between capillary suspension networks with and without nanoparticles. FINDINGS: Our observation suggests that nanoparticles induce the formation of thin liquid films on the surface of the microparticles, mitigating contact line pinning and promoting internal liquid exchange. Additionally, nanoparticles at microparticle contact regions further diminish Hertzian contact, enhancing the capacity for rearrangement. These effects enhance microparticle movement, narrowing the bridge size distribution.

Formation of Giant Unilamellar Vesicles Assisted by Fluorinated Nanoparticles.

In the quest to produce artificial cells, one key challenge that remains to be solved is the recreation of a complex cellular membrane. Among the existing models, giant unilamellar vesicles (GUVs) are particularly interesting due to their intrinsic compartmentalisation ability and their resemblance in size and shape to eukaryotic cells. Many techniques have been developed to produce GUVs all having inherent advantages and disadvantages. Here, the authors show that fluorinated silica nanoparticles (FNPs) used to form Pickering emulsions in a fluorinated oil can destabilise lipid nanosystems to template the formation of GUVs. This technique enables GUV production across a broad spectrum of buffer conditions, while preventing the leakage of the encapsulated components into the oil phase. Furthermore, a simple centrifugation process is sufficient for the release of the emulsion-trapped GUVs, bypassing the need to use emulsion-destabilising chemicals. With fluorescent FNPs and transmission electron microscopy, the authors confirm that FNPs are efficiently removed, producing contaminant-free GUVs. Further experiments assessing the lateral diffusion of lipids and unilamellarity of the GUVs demonstrate that they are comparable to GUVs produced via electroformation. Finally, the ability of incorporating transmembrane proteins is demonstrated, highlighting the potential of this method for the production of GUVs for artificial cell applications.

Stress-controlled shear flow alignment of collagen type I hydrogel systems.

Disease research and drug screening platforms require in vitro model systems with cellular cues resembling those of natural tissues. Fibrillar alignment, occurring naturally in extracellular matrices, is one of the crucial attributes in tissue development. Obtaining fiber alignment in 3D, in vitro remains an important challenge due to non-linear material characteristics. Here, we report a cell-compatible, shear stress-based method allowing to obtain 3D homogeneously aligned fibrillar collagen hydrogels. Controlling the shear-stress during gelation results in low strain rates, with negligible effects on the viability of embedded SH-SY5Y cells. Our approach offers reproducibility and tunability through a paradigm shift: The shear-stress initiation moment, being the critical optimization parameter in the process, is related to the modulus of the developing gel, whereas state of the art methods often rely on a predefined time to initiate the alignment procedure. After curing, the induced 3D alignment is maintained after the release of stress, with a linear relation between the total acquired strain and the fiber alignment. This method is generally applicable to 3D fibrillar materials and stress/pressure-controlled setups, making it a valuable addition to the fast-growing field of tissue engineering. STATEMENT OF SIGNIFICANCE: Controlling fiber alignment in vitro 3D hydrogels is crucial for developing physiologically relevant model systems. However, it remains challenging due to the non-linear material characteristics of fibrillar hydrogels, limiting the scalability and repeatability. Our approach tackles these challenges by utilizing a stress-controlled rheometer allowing us to monitor structural changes in situ and determine the optimal moment for applying a shear-stress inducing alignment. By careful parameter control, we infer the relationship between time, induced strain, alignment and biocompatibility. This tunable and reproducible method is both scalable and generally applicable to any fibrillar hydrogel, therefore, we believe it is useful for research investigating the link between matrix anisotropy and cell behavior in 3D systems, organ-on-chip technologies and drug research.

Glycyrrhizin-Based Hydrogels Accelerate Wound Healing of Normoglycemic and Diabetic Mouse Skin.

Efficient wound repair is crucial for mammalian survival. Healing of skin wounds is severely hampered in diabetic patients, resulting in chronic non-healing wounds that are difficult to treat. High-mobility group box 1 (HMGB1) is an important signaling molecule that is released during wounding, thereby delaying regenerative responses in the skin. Here, we show that dissolving glycyrrhizin, a potent HMGB1 inhibitor, in water results in the formation of a hydrogel with remarkable rheological properties. We demonstrate that these glycyrrhizin-based hydrogels accelerate cutaneous wound closure in normoglycemic and diabetic mice by influencing keratinocyte migration. To facilitate topical application of glycyrrhizin hydrogels on cutaneous wounds, several concentrations of glycyrrhizinic acid in water were tested for their rheological, structural, and biological properties. By varying the concentration of glycyrrhizin, these hydrogel properties can be readily tuned, enabling customized wound care.

Using an added liquid to suppress drying defects in hard particle coatings.

HYPOTHESIS: Lateral accumulation and film defects during drying of hard particle coatings is a common problem, typically solved using polymeric additives and surface active ingredients, which require further processing of the dried film. Capillary suspensions with their tunable physical properties, devoid of polymers, offer new pathways in producing uniform and defect free particulate coatings. EXPERIMENTS: We investigated the effect of small amounts of secondary liquid on the coating's drying behavior. Stress build-up and weight loss in a temperature and humidity controlled drying chamber were simultaneously measured. Changes in the coating's reflectance and height profile over time were related with the weight loss and stress curve. FINDINGS: Capillary suspensions dry uniformly without defects. Lateral drying is inhibited by the high yield stress, causing the coating to shrink to an even height. The bridges between particles prevent air invasion and extend the constant drying period. The liquid in the lower layers is transported to the interface via corner flow within surface pores, leading to a partially dry layer near the substrate while the pores above are still saturated. Using capillary suspensions for hard particle coatings results in more uniform, defect free films with better printing characteristics, rendering high additive content obsolete.

Apparatus for simultaneous stress and weight measurement during film drying in an environmentally controlled chamber.

The drying behavior of coatings is essential for the development of formulations in order to obtain reliable and defect free finishes. There are two major measures of interest: the development of the stress responsible for cracking and the drying rate that gives insight into the morphological structure. The cantilever deflection method is the predominant way of determining stresses under defined drying conditions such as temperature and humidity. However, both measures of interest are currently obtained using two different coatings when dried in a chamber or a single coating with simultaneous measurements that can only be dried under ambient conditions. In this paper, we present an apparatus design based on the cantilever deflection method that allows simultaneous measurement of the stress and drying rate in an environmentally controlled chamber.

Lightweight Porous Glass Composite Materials Based on Capillary Suspensions.

In this article, we present a simple, advanced method to produce lightweight tailor-made materials based on capillary suspensions that are made from locally bonded hollow glass spheres with a high total porosity in the range of 70% at apparent densities of 200 kg/m³, having a compressive strength of 0.6 MPa. The amount of added liquid and the particle surface treatment determine the network structure in the pastes and the resulting microstructure of the porous material in a straightforward manner. This structure has a strong impact on the porosity, pore size, and mechanical properties of the final body. The most promising porous materials were made of surface treated hollow glass spheres that create a sample-spanning network in the capillary state, where the added liquid wets the particles worse than the bulk fluid. These samples approach the density of natural balsa wood and they may find application in fields where either weight or structure are important, such as in insulation materials, filters, and membranes, as well as lightweight construction materials for automotive or aerospace engineering.