Comparison of Two Tricalcium Phosphate Varnishes and a Comparator Fluoride Varnish on Tubular Occlusion

Objectives The aim of the present study involved comparing the tubular occluding properties of three varnishes, two Clinpro™ varnishes and a Colgate® Duraphat® Varnish. Method Nine caries-free premolar dentine discs were prepared and treated with the three varnishes and subsequently observed under SEM at 0o and 90o angle to assess their ability to cover the dentine surface. The tubule occluding properties were measured using a modified Pashley cell hydraulic conductance model. A further 9 caries-free extracted molars were sectioned into 500μ thick dentine discs. The fluid flow rate was assessed after a) immersing the discs in 6% citric acid for 2 minutes, b) treating the dentine disc with the experimental varnishes and c) subjecting the treated discs to an acid challenge (6% citric acid for 2 minutes). Results SEM investigation showed uniform occlusion of the dentinal tubules, with varying depths of penetration. Hydraulic conductance tests showed no statistically significant differences in the fluid flow rate (expressed as percentages) when all the three varnishes were compared at different stages of treatment (p = 0.33). However, after subjecting the discs to an acid challenge, there was a statistically significant increase in the fluid flow rate with the Colgate® Duraphat® Varnish treated discs, whereas the ClinproTM White Varnish and Clinpro™ XT Varnish discs showed no statistically significant differences (p = 0.99 and p= 0.83 respectively). Conclusions All the tested varnishes (Colgate® Duraphat® Varnish, ClinproTM White Varnish and ClinproTM XT varnish) were effective in blocking the dentinal tubules as demonstrated in this in vitro study. However, the tricalcium phosphate varnishes (ClinproTM) were more resistant to an in vitro acid challenge compared to the sodium fluoride varnish (Colgate® Duraphat®)

Confocal laser scanning, scanning electron, and transmission electron microscopy investigation of Enterococcus faecalis biofilm degradation using passive and active sodium hypochlorite irrigation within a simulated root canal model

Root canal irrigation is an important adjunct to control microbial infection. The aim of this study was to investigate the effect of 2.5% (wt/vol) sodium hypochlorite (NaOCl) agitation on the removal, killing, and degradation of Enterococcus faecalis biofilm. A total of 45 root canal models were manufactured using 3D printing with each model comprising an 18 mm length simulated root canal of apical size 30 and taper 0.06. E. faecalis biofilms were grown on the apical 3 mm of the models for 10 days. A total of 60 s of 9 ml of 2.5% NaOCl irrigation using syringe and needle was performed, the irrigant was either left stagnant in the canal or agitated using manual (Gutta-percha), sonic, and ultrasonic methods for 30 s. Following irrigation, the residual biofilms were observed using confocal laser scanning, scanning electron, and transmission electron microscopy. The data were analyzed using one-way ANOVA with Dunnett post hoc tests at a level of significance p ≤ .05. Consequence of root canal irrigation indicate that the reduction in the amount of biofilm achieved with the active irrigation groups (manual, sonic, and ultrasonic) was significantly greater when compared with the passive and untreated groups (p < .05). Collectively, finding indicate that passive irrigation exhibited more residual biofilm on the model surface than irrigant agitated by manual or automated (sonic, ultrasonic) methods. Total biofilm degradation and nonviable cells were associated with the ultrasonic group

Quantitative nanohistological investigation of scleroderma: An atomic force microscopy-based approach to disease characterization

Scleroderma (or systemic sclerosis, SSc) is a disease caused by excess crosslinking of collagen. The skin stiffens and becomes painful, while internally, organ function can be compromised by the less elastic collagen. Diagnosis of SSc is often only possible in advanced cases by which treatment time is limited. A more detailed analysis of SSc may provide better future treatment options and information of disease progression. Recently, the histological stain picrosirius red showing collagen register has been combined with atomic force microscopy (AFM) to study SSc. Skin from healthy individuals and SSc patients was biopsied, stained and studied using AFM. By investigating the crosslinking of collagen at a smaller hierarchical stage, the effects of SSc were more pronounced. Changes in morphology and Young’s elastic modulus were observed and quantified; giving rise to a novel technique, we have termed “quantitative nanohistology”. An increase in nanoscale stiffness in the collagen for SSc compared with healthy individuals was seen by a significant increase in the Young’s modulus profile for the collagen. These markers of stiffer collagen in SSc are similar to the symptoms experienced by patients, giving additional hope that in the future, nanohistology using AFM can be readily applied as a clinical tool, providing detailed information of the state of collagen

Cell morphology as a design parameter in the bioengineering of cell-biomaterial surface interactions

Control of cell–surface interaction is necessary for biomaterial applications such as cell sheets, intelligent cell culture surfaces, or functional coatings. In this paper, we propose the emergent property of cell morphology as a design parameter in the bioengineering of cell–biomaterial surface interactions. Cell morphology measured through various parameters can indicate ideal candidates for these various applications thus reducing the time taken for the screening and development process. The hypothesis of this study is that there is an optimal cell morphology range for enhanced cell proliferation and migration on the surface of biomaterials. To test the hypothesis, primary porcine dermal fibroblasts (PDF, 3 biological replicates) were cultured on ten different surfaces comprising components of the natural extracellular matrix of tissues. Results suggested an optimal morphology with a cell aspect ratio (CAR) between 0.2 and 0.4 for both increased cell proliferation and migration. If the CAR was below 0.2 (very elongated cell), cell proliferation was increased whilst migration was reduced. A CAR of 0.4+ (rounded cell) favoured cell migration over proliferation. The screening process, when it comes to biomaterials is a long, repetitive, arduous but necessary event. This study highlights the beneficial use of testing the cell morphology on prospective prototypes, eliminating those that do not support an optimal cell shape. We believe that the research presented in this paper is important as we can help address this screening inefficiency through the use of the emergent property of cell morphology. Future work involves automating CAR quantification for high throughput screening of prototypes

Bioinspired Multifunctional Glass Surfaces through Regenerative Secondary Mask Lithography

Nature-inspired nanopatterning offers exciting multifunctionality spanning antireflectance and the ability to repel water/fog, oils, and bacteria; strongly dependent upon nanofeature size and morphology. However, such patterning in glass is notoriously difficult, paradoxically, due to the same outstanding chemical and thermal stability that make glass so attractive. Here, regenerative secondary mask lithography is introduced and exploited to enable customized glass nanopillars through dynamic nanoscale tunability of the side-wall profile and aspect ratio (>7). The method is simple and versatile, comprising just two steps. First, sub-wavelength scalable soft etch masks (55–350 nm) are generated through an example of block copolymer micelles or nanoimprinted photoresist. Second, their inherent durability problem is addressed by an innovative cyclic etching, when the original mask becomes embedded within a protective secondary organic mask, which is tuned and regenerated, permitting dynamic nanofeature profiling with etching selectivity >1:32. It is envisioned that such structuring in glass will facilitate fundamental studies and be useful for numerous practical applications—from displays to architectural windows. To showcase the potential, glass features are tailored to achieve excellent broadband omnidirectional antireflectivity, self-cleaning, and unique antibacterial activity toward Staphylococcus aureus

Atypical Mesenchymal Stromal Cell Responses to Topographic Modifications of Titanium Biomaterials Indicate Cytoskeletal- and Genetic Plasticity-Based Heterogeneity of Cells

Titanium (Ti) is widely used as a biomaterial for endosseous implants due to its relatively inert surface oxide layer that enables implanted devices the ability of assembling tissue reparative components that culminate in osseointegration. Topographic modifications in the form of micro- and nanoscaled structures significantly promote osseointegration and enhance the osteogenic differentiation of adult mesenchymal stromal cells (MSCs). While the biological mechanisms central to the differential responses of tissues and cells to Ti surface modifications remain unknown, adhesion and morphological adaptation are amongst the earliest events at the cell-biomaterial interface that are highly influenced by surface topography and profoundly impact the regulation of stem cell fate determination. This study correlated the effects of Ti topographic modifications on adhesion and morphological adaptation of human MSCs with phenotypic change. The results showed that modified Ti topographies precluded the adhesion of a subset of MSCs while incurring distinct morphological constraints on adherent cells. These effects anomalously corresponded with a differential expression of stem cell pluripotency and Wnt signalling-associated markers on both modified surfaces while additionally differing between hydrophobic and hydrophilic surface modifications—though extent of osteogenic differentiation induced by both modified topographies yielded similarly significant higher levels of cellular mineralisation in contrast to polished Ti. These results suggest that in the absence of deposited proteins and soluble factors, both modified topographies incur the selective adhesion of a subpopulation of progenitors with relatively higher cytoskeletal plasticity. While the presence of deposited proteins and soluble factors does not significantly affect adherence of cells, nanotopographic modifications enhance expression of pluripotency markers in proliferative conditions, which are conversely overridden by both modified topographies in osteogenic inductive conditions. Further deciphering the mechanisms underlying cellular selectivity and Ti topographic responsiveness will improve our understanding of stem cell heterogeneity and advance the potential of MSCs in regenerative medicine.</jats:p

Zinc and Strontium based Phosphate glass beads: a novel material for bone tissue engineering

Degradable phosphate-based glasses that contain strontium, zinc and calcium were investigated to examine its function as an osteoconductive material. Glass beads of the general formula of (P2O5)-(Na2O)-(TiO2)-(CaO)-(SrO) or (ZnO) were prepared by melt quench technique followed by milling and spheroidisation. Glass bead size distribution was initially measured by SEM. Then, some of these samples were immersed in deionized water to evaluate both the surface changes and measure the ion release rate, whereas other glass beads samples were incubated in culture media to determine pH changes. Furthermore, human osteoblast-like osteosarcoma cells MG-63 and human mesenchymal stem cells were seeded on the glass beads to determine its cytocompatibility via applying CCK assay, ALP assay and Ca assay. SEM images and fluorescence images of confocal microscopy were performed for the cellular studies. While mass degradation and ion release results displayed a significant increase with zinc and strontium incorporation within time, pH results showed an initial increase in pH followed by a decrease. Cellular studies emphasised that all formulations enhanced cellular proliferation. More specifically, ZnO5 and SrO17.5 displayed more promising results although they were insignificantly different from that of control (p&gt;0.05). This may suggest their applicability in hard tissue engineering

Impact of Carbamide Peroxide Whitening Agent on Dentinal Collagen

Carbamide peroxide (CP) is widely used as a tooth-whitening agent in self-administered tooth-bleaching products. In this study, the effects of 5% and 10% CP on dentinal collagen structure and chemical properties were evaluated in vitro. Thirty-five intact teeth were exposed to 2 whitening protocols (2 or 4 h daily) with either 5% or 10% CP gel for 1 wk. Shade changes before and after the whitening protocol were captured colorimetrically using a spectroshade. Collagen scaffold models and demineralized dentine disc samples were prepared and exposed to CP droplets (5% or 10%). Structural changes were investigated using electron microscopy. Finally, mineralized dentine disc samples were prepared postbleaching to assess chemical changes resulting from CP exposure in dentinal collagen using Raman spectroscopy. Results showed a difference in tooth shade when exposed to 5% and 10% CP whitening protocols, with a significantly (P ≤ 0.01) greater change reported for the 10% CP/4-h group. Imaging of the collagen scaffold model following exposure to CP showed a gelatinization process indicating that the free radical by-products from CP are able to disrupt the quaternary structure of noncrosslinked collagen. The most significant damage on the collagen scaffold was seen for the 10% CP exposure for 4 h. Imaging of the demineralized discs displayed the same glassy amorphous layer appearance as found in the collagen scaffold. Raman spectra of the mineralized dentine discs showed a significant decrease (P ≤ 0.01) in the integrated area of amide I and amide III values in the 4 test groups following CP application. Amide I was more affected as both the exposure time and concentration of CP increased. Despite the claimed safety of whitening agents, this in vitro study concludes that even low concentrations of CP result in a deleterious change in dentinal collagen

Investigations into in situ Enterococcus faecalis biofilm removal by passive and active sodium hypochlorite irrigation delivered into the lateral canal of a simulated root canal model

Aim To investigate in situ Enterococcus faecalis biofilm removal from the lateral canal of a simulated root canal system using passive or active irrigation protocols. Methodology Root canal models (n = 43) were manufactured from transparent resin materials using 3D-printing. Each canal was created with an 18 mm length, apical size 30, a .06 taper, and a lateral canal of 3 mm length, 0.3 mm diameter. Biofilms were grown on the lateral canal and apical 3 mm of the main canal for 10 days. Biofilm of three models was examined using SEM. The other forty models were divided to four groups (n = 10). The models were observed under a fluorescence microscope. Following 60 s of 9 mL of 2.5% NaOCl irrigation using syringe and needle, the irrigant was either left stagnant in the canal or activated using gutta-percha, sonic or ultrasonic methods for 30 s. Images were then captured every second using an external camera. The residual biofilm percentages were measured using image analysis software. The data were analyzed using generalized linear mixed models. A significance level of 0.05 was used throughout. Results The greatest level of biofilm removal was with ultrasonic agitation (66.76%) followed by sonic (45.49%), manual agitation (43.97%), and passive irrigation groups (38.67%) respectively. The differences were significant between the residual biofilm in the passive irrigation and both sonic & ultrasonic groups (P = 0.001). Conclusion Agitation resulted in better penetration of the 2.5% NaOCl into the lateral canal of an artificial root canal model. Ultrasonic agitation of NaOCl improved the removal of biofilm

Vitamin D and Its Role During Pregnancy in Attaining Optimal Health of Mother and Fetus

Despite its discovery a hundred years ago, vitamin D has emerged as one of the most controversial nutrients and prohormones of the 21st century. Its role in calcium metabolism and bone health is undisputed but its role in immune function and long-term health is debated. There are clear indicators from in vitro and animal in vivo studies that point to vitamin D’s indisputable role in both innate and adaptive immunity; however, the translation of these findings to clinical practice, including the care of the pregnant woman, has not occurred. Until recently, there has been a paucity of data from randomized controlled trials to establish clear cut beneficial effects of vitamin D supplementation during pregnancy. An overview of vitamin metabolism, states of deficiency, and the results of recent clinical trials conducted in the U.S. are presented with an emphasis on what is known and what questions remain to be answered