Implications of a cheliceral axial duplication in Tetragnatha versicolor (Araneae: Tetragnathidae) for arachnid deuterocerebral appendage development.

The homology of the arachnid chelicera with respect to other head appendages in Panarthropoda has long been debated. Gene expression data and the re-interpretation of early transitional fossils have supported the homology of the deutocerebrum and its associated appendages, implying a homology between primary antennae (mandibulates), chelicerae (euchelicerates), and chelifores (sea spiders). Nevertheless, comparatively little is known about the mechanistic basis of proximo-distal (PD) axis induction in chelicerates, much less the basis for cheliceral fate specification. Here, we describe a new cheliceral teratology in the spider Tetragnatha versicolor Walckenaer, 1841, which consists on a duplication of the PD axis of the left chelicera associated with a terminal secondary schistomely on the fang of the lower axis. This duplication offers clues as to potential shared mechanisms of PD axis formation in the chelicera. We review the state of knowledge on PD axis induction mechanisms in arthropods and identify elements of gene regulatory networks that are key for future functional experiments of appendage development in non-insect model systems. Such investigations would allow a better understanding of PD axis induction of modified and poorly studied arthropod limbs (e.g., chelicerae, chelifores, and ovigers).

Lubricating properties of chewing stimulated whole saliva from patients suffering from xerostomia.

OBJECTIVES: The study aimed to quantify the lubricating properties of chewing stimulated whole saliva from healthy controls (n = 22), from patients suffering from primary Sjögren's syndrome (n = 37) and from patients undergoing head-and-neck radiotherapy (n = 34). MATERIALS AND METHODS: All participants had to complete the Xerostomia Inventory questionnaire to score dry mouth sensation. Lubrication was measured using an ex vivo tongue-enamel friction system in terms of Relief and Relief period. MUC5b and total protein concentrations of the saliva samples were measured by an enzyme-linked immunosorbent assay and a bicinchoninic acid assay, respectively. RESULTS: Relief of Sjögren's patients' saliva and post-irradiation patients' saliva was similar compared with healthy controls, but saliva from post-irradiation patients lubricated significantly better than saliva from Sjögren's patients. The Relief period was similar between the three groups. The Relief and Relief period were higher for saliva samples post-irradiation compared to pre-irradiation. MUC5b and total protein concentrations were comparable in all groups. MUC5b and total protein output were significantly lower in patients subjected to radiotherapy compared to saliva from healthy controls and pre-irradiation patients. MUC5b concentrations positively correlated with lubricating properties of post-irradiation patient saliva. CONCLUSIONS: The lubricating properties of patient saliva were not any worse than healthy controls. Lower flow rate leads to lower availability of saliva in the oral cavity and decreases the overall output of protein and MUC5b, which might result in an insufficient replenishing of the mucosal salivary film. CLINICAL RELEVANCE: An insufficient replenishing might underlie the sensation of a dry mouth and loss of oral function.

Adipose Tissue-Derived Stromal Cells Alter the Mechanical Stability and Viscoelastic Properties of Gelatine Methacryloyl Hydrogels.

The extracellular matrix provides mechanical cues to cells within it, not just in terms of stiffness (elasticity) but also time-dependent responses to deformation (viscoelasticity). In this work, we determined the viscoelastic transformation of gelatine methacryloyl (GelMA) hydrogels caused by adipose tissue-derived stromal cells (ASCs) through mathematical modelling. GelMA-ASCs combination is of interest to model stem cell-driven repair and to understand cell-biomaterial interactions in 3D environments. Immortalised human ASCs were embedded in 5%, 10%, and 15% (w/v) GelMA hydrogels and evaluated for 14 d. GelMA had a concentration-dependent increase in stiffness, but cells decreased this stiffness over time, across concentrations. Viscoelastic changes in terms of stress relaxation increased progressively in 5% GelMA, while mathematical Maxwell analysis showed that the relative importance (Ri) of the fastest Maxwell elements increased proportionally. The 10% GelMA only showed differences at 7 d. In contrast, ASCs in 15% GelMA caused slower stress relaxation, increasing the Ri of the slowest Maxwell element. We conclude that GelMA concentration influenced the stiffness and number of Maxwell elements. ASCs changed the percentage stress relaxation and Ri of Maxwell elements transforming hydrogel viscoelasticity into a more fluid environment over time. Overall, 5% GelMA induced the most favourable ASC response.

Correction to: Dry mouth: saliva substitutes which adsorb and modify existing salivary condition films improve oral lubrication.

After publication of this paper, the authors observed that that figure 6 appears before figure 5.

Dry mouth: saliva substitutes which adsorb and modify existing salivary condition films improve oral lubrication.

OBJECTIVES: The aims of this study are to assess different saliva substitutes for their efficacy to lubricate the oral cavity, and to relate this oral lubrication to the ability of saliva substitutes to adsorb on and change the structure of the existing salivary conditioning film (SCF). MATERIALS AND METHODS: Quartz crystal microbalance with dissipation was used to study the capability of saliva substitutes to interact with natural SCF and the ability to change the secondary SCF (S-SCF). A tongue-enamel friction system mimicking xerostomic conditions was used to assess the relief and relief period expected from these substitutes under set circumstances. RESULTS: Saliva Orthana spray, Biotène spray and Gum Hydral gel had an immediate effect on a SCF, increasing its structural softness. BioXtra gel, Biotène gel, Gum Hydral gel and Glandosane spray changed the S-SCF by increasing salivary protein adsorption, while others showed no sign of interaction. With respect to relief, only 2 out of the 16 saliva substitutes tested (Saliva Orthana spray and Gum Hydral gel) performed better than water. Overall, relief period correlated positively to structural softness change, whereas a positive correlation was seen between relief and mass adsorption. CONCLUSIONS: The majority of saliva substitutes did not adsorb on the SCF, thus did not enhance lubrication. Only saliva substitutes containing carrageenan, carboxymethylcellulose, pig gastric mucin, xanthan gum and carbomer performed better in enhancing oral lubrication. CLINICAL RELEVANCE: This objective assessment will help clinicians and patients make better choice of saliva substitutes. This study provides a scientific basis for future improvement in saliva substitutes.

Expression and function of spineless orthologs correlate with distal deutocerebral appendage morphology across Arthropoda.

The deutocerebral (second) head segment is putatively homologous across Arthropoda, in spite of remarkable disparity of form and function of deutocerebral appendages. In Mandibulata this segment bears a pair of sensory antennae, whereas in Chelicerata the same segment bears a pair of feeding appendages called chelicerae. Part of the evidence for the homology of deutocerebral appendages is the conserved function of homothorax (hth), which has been shown to specify antennal or cheliceral fate in the absence of Hox signaling, in both mandibulate and chelicerate exemplars. However, the genetic basis for the morphological disparity of antenna and chelicera is not understood. To test whether downstream targets of hth have diverged in a lineage-specific manner, we examined the evolution of the function and expression of spineless (ss), which in two holometabolous insects is known to act as a hth target and distal antennal determinant. Toward expanding phylogenetic representation of gene expression data, here we show that strong expression of ss is observed in developing antennae of a hemimetabolous insect, a centipede, and an amphipod crustacean. By contrast, ss orthologs are not expressed throughout the cheliceral limb buds of spiders or harvestmen during developmental stages when appendage fate is specified. RNA interference-mediated knockdown of ss in Oncopeltus fasciatus, which bears a simple plesiomorphic antenna, resulted in homeotic distal antenna-to-leg transformation, comparable to data from holometabolous insect counterparts. Knockdown of hth in Oncopeltus fasciatus abrogated ss expression, suggesting conservation of upstream regulation. These data suggest that ss may be a flagellar (distal antennal) determinant more broadly, and that this function was acquired at the base of Mandibulata.

Generation of protective immunity against Orientia tsutsugamushi infection by immunization with a zinc oxide nanoparticle combined with ScaA antigen.

BACKGROUND: Zinc oxide nanoparticle (ZNP) has been applied in various biomedical fields. Here, we investigated the usage of ZNP as an antigen carrier for vaccine development by combining a high affinity peptide to ZNP. RESULTS: A novel zinc oxide-binding peptide (ZBP), FPYPGGDA, with high affinity to ZNP (K a  = 2.26 × 106 M-1) was isolated from a random peptide library and fused with a bacterial antigen, ScaA of Orientia tsutsugamushi, the causative agent of scrub typhus. The ZNP/ZBP-ScaA complex was efficiently phagocytosed by a dendritic cell line, DC2.4, in vitro and significantly enhanced anti-ScaA antibody responses in vivo compared to control groups. In addition, immunization with the ZNP/ZBP-ScaA complex promoted the generation of IFN-γ-secreting T cells in an antigen-dependent manner. Finally, we observed that ZNP/ZBP-ScaA immunization provided protective immunity against lethal challenge of O. tsutsugamushi, indicating that ZNP can be used as a potent adjuvant when complexed with ZBP-conjugated antigen. CONCLUSIONS: ZNPs possess good adjuvant potential as a vaccine carrier when combined with an antigen having a high affinity to ZNP. When complexed with ZBP-ScaA antigen, ZNPs could induce strong antibody responses as well as protective immunity against lethal challenges of O. tsutsugamushi. Therefore, application of ZNPs combined with a specific soluble antigen could be a promising strategy as a novel vaccine carrier system.

Phylogenomic interrogation of arachnida reveals systemic conflicts in phylogenetic signal.

Chelicerata represents one of the oldest groups of arthropods, with a fossil record extending to the Cambrian, and is sister group to the remaining extant arthropods, the mandibulates. Attempts to resolve the internal phylogeny of chelicerates have achieved little consensus, due to marked discord in both morphological and molecular hypotheses of chelicerate phylogeny. The monophyly of Arachnida, the terrestrial chelicerates, is generally accepted, but has garnered little support from molecular data, which have been limited either in breadth of taxonomic sampling or in depth of sequencing. To address the internal phylogeny of this group, we employed a phylogenomic approach, generating transcriptomic data for 17 species in combination with existing data, including two complete genomes. We analyzed multiple data sets containing up to 1,235,912 sites across 3,644 loci, using alternative approaches to optimization of matrix composition. Here, we show that phylogenetic signal for the monophyly of Arachnida is restricted to the 500 slowest-evolving genes in the data set. Accelerated evolutionary rates in Acariformes, Pseudoscorpiones, and Parasitiformes potentially engender long-branch attraction artifacts, yielding nonmonophyly of Arachnida with increasing support upon incrementing the number of concatenated genes. Mutually exclusive hypotheses are supported by locus groups of variable evolutionary rate, revealing significant conflicts in phylogenetic signal. Analyses of gene-tree discordance indicate marked incongruence in relationships among chelicerate orders, whereas derived relationships are demonstrably robust. Consistently recovered and supported relationships include the monophyly of Chelicerata, Euchelicerata, Tetrapulmonata, and all orders represented by multiple terminals. Relationships supported by subsets of slow-evolving genes include Ricinulei + Solifugae; a clade comprised of Ricinulei, Opiliones, and Solifugae; and a clade comprised of Tetrapulmonata, Scorpiones, and Pseudoscorpiones. We demonstrate that outgroup selection without regard for branch length distribution exacerbates long-branch attraction artifacts and does not mitigate gene-tree discordance, regardless of high gene representation for outgroups that are model organisms. Arachnopulmonata (new name) is proposed for the clade comprising Scorpiones + Tetrapulmonata (previously named Pulmonata).

A conserved genetic mechanism specifies deutocerebral appendage identity in insects and arachnids.

The segmental architecture of the arthropod head is one of the most controversial topics in the evolutionary developmental biology of arthropods. The deutocerebral (second) segment of the head is putatively homologous across Arthropoda, as inferred from the segmental distribution of the tripartite brain and the absence of Hox gene expression of this anterior-most, appendage-bearing segment. While this homology statement implies a putative common mechanism for differentiation of deutocerebral appendages across arthropods, experimental data for deutocerebral appendage fate specification are limited to winged insects. Mandibulates (hexapods, crustaceans and myriapods) bear a characteristic pair of antennae on the deutocerebral segment, whereas chelicerates (e.g. spiders, scorpions, harvestmen) bear the eponymous chelicerae. In such hexapods as the fruit fly, Drosophila melanogaster, and the cricket, Gryllus bimaculatus, cephalic appendages are differentiated from the thoracic appendages (legs) by the activity of the appendage patterning gene homothorax (hth). Here we show that embryonic RNA interference against hth in the harvestman Phalangium opilio results in homeonotic chelicera-to-leg transformations, and also in some cases pedipalp-to-leg transformations. In more strongly affected embryos, adjacent appendages undergo fusion and/or truncation, and legs display proximal defects, suggesting conservation of additional functions of hth in patterning the antero-posterior and proximo-distal appendage axes. Expression signal of anterior Hox genes labial, proboscipedia and Deformed is diminished, but not absent, in hth RNAi embryos, consistent with results previously obtained with the insect G. bimaculatus. Our results substantiate a deep homology across arthropods of the mechanism whereby cephalic appendages are differentiated from locomotory appendages.

Changes in lens stiffness due to capsular opacification in accommodative lens refilling.

Accommodation may be restored to presbyopic lenses by refilling the lens capsular bag with a soft polymer. After this accommodative lens refilling prevention of capsular opacification is a requirement, since capsular opacification leads to a decreased clarity of the refilled lens. It has been hypothesized that capsular fibrosis causing the capsular opacification results in increased stiffness of the lens capsular bag, therewith contributing to a decrease in accommodative amplitude of the lens. However, the change in viscoelastic properties of refilled lenses due to capsular fibrosis has never been measured directly. In this study we examined natural lenses from enucleated porcine eyes and refilled lenses directly after refilling and after three months of culturing, when capsular fibrosis had developed, and determined their viscoelastic properties with a low load compression tester. Control refilled lenses were included in which capsular opacification was prevented by treatment with actinomycin D. We related lens stiffening to the degree of capsular opacification, as derived from the microscopic images taken with a confocal laser scanning microscope. Overall, the refilled lenses directly after refilling were softer than refilled lenses after three months of culturing, and refilled lenses treated with actinomycin D were softer compared with untreated refilled lenses. The degree of capsular opacification as assessed by microscopy corresponds to an increase in lens stiffness. This indicates that the viscoelastic properties of the refilled lens are influenced by capsular fibrosis and modulated by treatment of the lens epithelium. In conclusion, this study shows that the development of capsular fibrosis negatively affects the viscoelastic properties of isolated, cultured refilled lenses.

Visualization of microbiological processes underlying stress relaxation in Pseudomonas aeruginosa biofilms.

Bacterial biofilms relieve themselves from external stresses through internal rearrangement, as mathematically modeled in many studies, but never microscopically visualized for their underlying microbiological processes. The aim of this study was to visualize rearrangement processes occurring in mechanically deformed biofilms using confocal-laser-scanning-microscopy after SYTO9 (green-fluorescent) and calcofluor-white (blue-fluorescent) staining to visualize bacteria and extracellular-polymeric matrix substances, respectively. We apply 20% uniaxial deformation to Pseudomonas aeruginosa biofilms and fix deformed biofilms prior to staining, after allowing different time-periods for relaxation. Two isogenic P. aeruginosa strains with different abilities to produce extracellular polymeric substances (EPS) were used. By confocal-laser-scanning-microscopy all biofilms showed intensity distributions for fluorescence from which rearrangement of EPS and bacteria in deformed biofilms were derived. For the P. aeruginosa strain producing EPS, bacteria could not find new, stable positions within 100 s after deformation, while EPS moved toward deeper layers within 20 s. Bacterial rearrangement was not seen in P. aeruginosa biofilms deficient in production of EPS. Thus, EPS is required to stimulate bacterial rearrangement in mechanically deformed biofilms within the time-scale of our experiments, and the mere presence of water is insufficient to induce bacterial movement, likely due to its looser association with the bacteria.

Presence of single nucleotide polymorphisms in transforming growth factor ß and insulin-like growth factor 1 in class II malocclusions due to retrognathic mandible.

AIM: The aim of this study was to evaluate specific single nucleotide polymorphisms (SNP) of transforming growth factor-beta (TGF-ß) (rs1800469) and insulin-like growth factor-1 (IGF-1) (rs17032362) genes in Class II individuals with a normal maxilla and retrognathic (short) mandible.

Influence of bottled salad dressings on the development of enamel erosion in the presence or absence of salivary pellicle - An in vitro study.

Background: Acidic beverages are believed to elevate the risk of enamel surface erosion. In addition to the intake of soft drinks, the increased consumption of salad dressings has been linked to a higher prevalence of dental erosion. Therefore, the current study aimed to investigate the influence of bottled salad dressings on the development of enamel erosion in the presence or absence of pellicle through in vitro experiment. Methods: Preliminary pH and calcium analyses of solutions were performed. Highest pH and calcium content was found for sandwich spread i.e., 4.69 and 55.4 mg/100 g grams, respectively. Eighty tooth specimens (measuring 4 × 4 × 3 mm) were prepared from extracted human premolars and randomly assigned to four groups (group 1: orange juice; group 2: eggless plain mayonnaise; group 3: sandwich spread; and group 4: thousand island dressing) with 20 samples in each group. Ten tooth specimens from each group were immersed in 20 ml of the respective solutions for 5 min (control group). The remaining ten tooth specimens from each group were submerged in 5 mL saliva vials for 3 min to facilitate salivary pellicle formation before being immersed in their respective solutions for 5 min (saliva-covered group). Pre and post-experimental assessments of enamel roughness and hardness were conducted using a surface roughness tester and Knoop Hardness indenter, respectively. Results: Overall, enamel roughness was notably elevated in the control group, with the eggless plain mayonnaise (0.52 ± 0.38) and thousand island dressing groups (0.57 ± 0.29) showing a significant increase in surface roughness post-test (p = 0.05). Nevertheless, there was no significant difference in the enamel roughness between the groups. On the other hand, regardless of the presence/absence of the salivary pellicle, a marked decrease in enamel hardness was observed among all groups except for group 3 (sandwich spread) with a mean score of 311.5 ± 82.6 (p < 0.05). Conclusion: A significant increase in surface roughness and reduction in enamel hardness was observed with salad dressings. However, in vitro formed salivary pellicle showed a protective effect against tooth erosion.

Effects of natural antimicrobial compounds propolis and copaiba on periodontal ligament fibroblasts, molecular docking, and in vivo study in Galleria mellonella.

Root canal treatment addresses infectious processes that require control. Occasionally, the radicular pulp is vital and inflamed, presenting a superficial infection. To preserve pulpal remnants, conservative procedures have gained favor, employing anti-inflammatory medications. This study investigated the effects of propolis (PRO), and copaiba oil-resin (COR) associated with hydrocortisone (H) and compared their impact to that of Otosporin® concerning cytotoxic and genotoxic activity, cytokine detection, and toxicity in the Galleria mellonella model. Human periodontal ligament fibroblasts (PDLFs) were exposed to drug concentrations and evaluated by the MTT assay. Associations were tested from concentrations that did not compromise cell density. Genotoxicity was evaluated through micronucleus counting, while cytokines IL-6 and TGF-ß1 were detected in the cell supernatant using ELISA. Molecular docking simulations were conducted, considering the major compounds identified in PRO, COR, and H. Increasing concentrations of PRO and COR were assessed for acute toxicity in Galleria mellonella model. Cellular assays were analyzed using one-way ANOVA followed by Tukey tests, while larval survivals were evaluated using the Log-rank (Mantel-Cox) test (α = 0.05). PRO and COR promoted PDLFs proliferation, even in conjunction with H. No changes in cell metabolism were observed concerning cytokine levels. The tested materials induce the release of AT1R, proliferating the PDFLs through interactions. PRO and COR had low toxicity in larvae, suggesting safety at tested levels. These findings endorse the potential of PRO and COR in endodontics and present promising applications across medical domains, such as preventive strategies in inflammation, shedding light on their potential development into commercially available drugs.

Boundary lubrication by brushed salivary conditioning films and their degree of glycosylation.

OBJECTIVES: Toothbrushing, though aimed at biofilm removal, also affects the lubricative function of adsorbed salivary conditioning films (SCFs). Different modes of brushing (manual, powered, rotary-oscillatory or sonically driven) influence the SCF in different ways. Our objectives were to compare boundary lubrication of SCFs after different modes of brushing and to explain their lubrication on the basis of their roughness, dehydrated layer thickness, and degree of glycosylation. A pilot study was performed to relate in vitro lubrication with mouthfeel in human volunteers. MATERIALS AND METHODS: Coefficient of friction (COF) on 16-h-old SCFs after manual, rotary-oscillatory, and sonically driven brushing was measured using colloidal probe atomic force microscopy (AFM). AFM was also used to assess the roughness of SCFs prior to and after brushing. Dehydrated layer thicknesses and glycosylation of the SCFs were determined using X-ray photoelectron spectroscopy. Mouthfeel after manual and both modes of powered brushing were evaluated employing a split-mouth design. RESULTS: Compared with unbrushed and manually or sonically driven brushed SCFs, powered rotary-oscillatory brushing leads to deglycosylation of the SCF, loss of thickness, and a rougher film. Concurrently, the COF of a powered rotary-oscillatory brushed SCF increased. Volunteers reported a slightly preferred mouthfeel after sonic brushing as compared to powered rotating-oscillating brushing. CONCLUSION: Deglycosylation and roughness increase the COF on SCFs. CLINICAL RELEVANCE: Powered rotary-oscillatory brushing can deglycosylate a SCF, leading to a rougher film surface as compared with manual and sonic brushing, decreasing the lubricative function of the SCF. This is consistent with clinical mouthfeel evaluation after different modes of brushing.

Emerging approaches of neural regeneration using physical stimulations solely or coupled with smart piezoelectric nano-biomaterials.

Neural regeneration is a challenging venture as it is limited by various intrinsic physiological parameters such as the presence of biomolecules like Nogo-A, Ephrin-B3 and Neurocan, that inhibit Central nervous system (CNS) regeneration, and the absence of conducive factors such as ATF3, Sox2 and GAP-43, that promote the neuronal differentiation and regeneration. The design of an effective strategy for neuronal repair or regeneration is a daunting task as neural cells are responsive to a very narrow window of the conductive cellular microenvironment. It requires specific inductive signals and chemical cues from neighbouring cells that can trigger the process of regeneration or repair. It is this complexity that adds to the plight in the scenarios of patients confronted with trauma resulting in spinal cord injury (SCI) or traumatic brain injury (TBI). SCI or TBI may cause temporary or permanent locomotory disorders in patients, affecting the quality of their lives. The regenerative potential of neural cells in the CNS is comparatively lesser than that of peripheral nervous system (PNS). Also, the activation and migration of astrocytes to the injury site causes glial scar, thus hindering further repair process, especially in CNS injuries. Therefore, an effective strategy for stimulating neuritic branching and growth can be a solution to the problem. This review discusses the various facets of strategies that have been adopted to understand and improve the progress of neural tissue engineering for treating the conditions like SCI and TBI. This review also provides an insight regarding the influence of various nano-topographical cues on neuronal cell behavior, the importance of inherent piezoelectric properties in biological systems, various forms of physical stimulation methods that can drive the process of neuritic outgrowth, and finally concludes with the elucidations of advances in development of various biomaterials that have been found effective in achieving enhancement in neuronal physiological properties. It also shares some opinions as perspectives that may help in the further advancement of this field.

Lectin-Functionalized Polyethylene Glycol for Relief of Mucosal Dryness.

The importance of lubrication between oral surfaces provided by the salivary film is most acutely apparent when it is disrupted, a prevalent consequence of salivary gland hypofunction experienced with aging, a symptom of certain diseases, or a side effect of some medical interventions. Sufferers report difficulty with speech and oral food processing and collectively is detrimental to quality of life. Polyethylene glycol (PEG) is widely employed as a successful biocompatible boundary lubricant in engineering and biomedical applications. It is hypothesized that the immobilization of PEG to biological materials such as oral epithelial cells and tissue can mimic the salivary film and provide durable relief from the symptoms of mucosal dryness. To do so, PEG is functionalized with a sugar binding lectin (wheat germ agglutinin) to enhance epithelial adhesion through lectin-sugar interactions. Retention and lubricity are characterized on an ex vivo oral tissue tribology rig. WGA-PEG coats and retains on mucin films, oral epithelial cells, and porcine tongue tissue, and offers sustained reduction in coefficient of friction (COF). WGA-PEG could be developed into a useful topical treatment for reducing oral friction and the perception of dry mouth.

The Vehicles of Calcium Hydroxide Pastes Interfere with Antimicrobial Effect, Biofilm Polysaccharidic Matrix, and Pastes' Physicochemical Properties.

The objective of the present study was to investigate the pH, volumetric alteration, antimicrobial action, and effect on biofilm matrix polysaccharides of calcium hydroxide (CH) pastes with different vehicles available in endodontics: CH + propylene glycol (CHP), UltraCal XS®, Metapaste®, and Metapex®. The pH was analyzed at different time intervals using a pH meter. For volumetric alteration, a microtomographic assay was performed before and after immersion in water. Enterococcus faecalis was chosen for microbiological tests. The bacterial viability and extracellular matrix were quantified with direct contact evaluation (dentin blocks) and at the intratubular level (dentin cylinders) using LIVE/DEAD BacLight and Calcofluor White dyes via confocal laser scanning microscopy (CLSM). Kruskal-Wallis and Dunn's tests were used to analyze pH and direct contact assays, while one-way ANOVA and Tukey tests were used to analyze volumetric alteration and intratubular decontamination (α = 0.05). Higher pH values were obtained during the initial days. Volumetric alterations were similar in all groups. Lower bacterial viability was obtained for dentin blocks and cylinders when CH pastes were used. UltraCal XS and Metapex had lower values for the extracellular matrix. The pH of all CH pastes decreased with time and did not promote medium alkalization for up to 30 days. CH paste can reduce bacterial viability through direct contact and at an intratubular level; however, UltraCal XS and Metapex are involved with lower volumes of extracellular matrices.

Influence of fluoride-detergent combinations on the visco-elasticity of adsorbed salivary protein films.

The visco-elasticity of salivary-protein films is related to mouthfeel, lubrication, biofilm formation, and protection against erosion and is influenced by the adsorption of toothpaste components. The thickness and the visco-elasticity of hydrated films (determined using a quartz crystal microbalance) of 2-h-old in vitro-adsorbed salivary-protein films were 43.5 nm and 9.4 MHz, respectively, whereas the dehydrated thickness, measured using X-ray photoelectron spectroscopy, was 2.4 nm. Treatment with toothpaste slurries decreased the thickness of the film, depending on the fluoride-detergent combination involved. Secondary exposure to saliva resulted in a regained thickness of the film to a level similar to its original thickness; however, no association was found between the thickness of hydrated and dehydrated films, indicating differences in film structure. Treatment with stannous fluoride/sodium lauryl sulphate (SnF(2)/SLS)-containing toothpaste slurries yielded a strong, immediate two-fold increase in characteristic film frequency (f(c)) with respect to untreated films, indicating cross-linking in adsorbed salivary-protein films by Sn(2+) that was absent when SLS was replaced with sodium hexametaphosphate (NaHMP). Secondary exposure to saliva of films treated with SnF(2) caused a strong, six-fold increase in f(c) compared with primary salivary-protein films, regardless of whether SLS or NaHMP was the detergent. This suggests that ionized stannous is not directly available for cross-linking in combination with highly negatively charged NaHMP, but becomes slowly available after initial treatment to cause cross-linking during secondary exposure to saliva.

Cartilage lamina splendens inspired nanostructured coating for biomaterial lubrication.

Biomaterials that are used in biological systems, such as polycarbonate urethane (PCU) knee joint implants and contact lenses, generally lack lubrication. This limits their integration with the body and impedes their function. Here, we propose a nanostructured film based on hydrophilic polysaccharide hyaluronic acid conjugated with dopamine (HADN) and zwitterionic reduced glutathione (Glu), which forms a composite coating (HADN-Glu) to enhance the lubrication between cartilage and PCU. HADN was synthesized by carbodiimide chemistry between hyaluronic acid and dopamine and deposited on PCU surface under mild oxidative conditions. Then, zwitterionic peptide-reduced glutathione was bioconjugated to HADN, forming a lubrication film. Analysis based on X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and wettability indicated that HADN and Glu had grafted successfully onto the PCU surface. Measurements of the coefficient of friction (COF), friction energy dissipation and cartilage roughness indicated that cartilage was effectively protected by the high lubrication of HADN-Glu. Both at low and high applied loads, this effect was likely due to the enhanced boundary lubrication enabled by HADN-Glu on the PCU surface. Moreover, HADN-Glu is highly biocompatible with chondrocyte cells, suggesting that this film will benefit the design of implants where lubrication is needed.