Elucidation of the basic mechanisms underlying human disease pathogenesis depends on the findings afforded to us through in vivo and in vitro approaches. While there are inherent limitations in any model system, 2D in vitro culture systems tend to be particularly restricted due to their static nature. Here, we adapted a flow-based hollow-fiber cartridge system to better understand the cellular influences of human retinal microvascular endothelial cells and mouse-derived neutrophils under high glucose conditions similar to those observed in diabetes. Analyses by western blot and flow cytometry indicate that pro-inflammatory molecules known to be associated with the pathogenesis of diabetic retinopathy were significantly elevated following high glucose exposure, including VEGF, ICAM-1, and ROS. Changes in mitochondrial potential were also observed. Further, we demonstrate that this innovative system allows for cross-species co-culture as well as long-term culturing conditions. This in vitro modeling system not only mimics the retinal microvasculature, it also allows for the examination of cellular interactions and mechanisms that contribute to diabetic retinopathy, a visually debilitating complication of diabetes.

Advanced platelet-rich fibrin (A-PRF) provides the scaffold and growth factors necessary for stem cells to proliferate and differentiate in successful regenerative endodontic procedures. This study investigates the release of transforming growth factor-β1 (TGF-β1), platelet-derived growth factor (PDGF), and vascular endothelial growth factor (VEGF) from A-PRF in cell culture media in the presence and absence of mineral trioxide aggregate (MTA) or Biodentine. Additionally, this research assesses the viability and migration of stem cells of the apical papilla (SCAP) in previously conditioned media. A-PRF obtained from 14 participants were incubated for 7 days in cell culture media alone or via layering with MTA or Biodentine discs and the release of selected growth factors in the media was evaluated using ELISA. The viability of SCAP grown in conditioned media was measured using the CCK8 assay, while SCAP migration was assessed via a transwell assay by counting migrated cells. The release of TGF-β1, PDGF, and VEGF was significantly higher in media with A-PRF alone than in the presence of either calcium-based silicate material (p < 0.05), which showed no difference from the no-A-PRF control (p < 0.05). None of the tested growth factors released in the A-PRF-conditioned media correlated with clot weight. A-PRF-conditioned media, both with and without calcium-based silicate materials, did not impact SCAP viability and migration (p > 0.05). This study shows that SCAP behavior is not impacted by the decrease in growth factor released in the presence of calcium-based silicate materials and that their role in REPs warrants further investigation.

This study investigated the influence of incorporating Biosilicate^® on the physico-mechanical and biological properties of glass ionomer cement (GIC). This bioactive glass ceramic (23.75% Na₂O, 23.75% CaO, 48.5% SiO₂, and 4% P₂O₅) was incorporated by weight (5%, 10%, or 15%) into commercially available GICs (Maxxion R and Fuji IX GP). Surface characterization was made by SEM (n = 3), EDS (n = 3), and FTIR (n = 1). The setting and working (S/W time) times (n = 3) and compressive strength (CS) were analyzed (n = 10) according to ISO 9917-1:2007. The ion release (n = 6) was determined and quantified by ICP OES and by UV-Vis for Ca, Na, Al, Si, P, and F. To verify cell cytotoxicity, stem cells from the apical papilla (SCAP) were exposed to eluates (n = 3, at a ratio of 1.8 cm²/mL) and analyzed 24 h post-exposure. Antimicrobial activity against Streptococcus mutans (ATCC 25175, NCTC 10449) was analyzed by direct contact for 2 h (n = 5). The data were submitted for normality and lognormality testing. One-way ANOVA and Tukey’s test were applied for the working and setting time, compressive strength, and ion release data. Data from cytotoxicity and antimicrobial activity were submitted for Kruskal–Wallis’ testing and Dunn’s post hoc test (α = 0.05). Among all experimental groups, only those with 5% (wt) of Biosilicate^® showed better surface quality. Only M5% showed a comparable W/S time to the original material (p = 0.7254 and p = 0.5912). CS was maintained for all Maxxion R groups (p > 0.0001) and declined for Fuji IX experimental groups (p < 0.0001). The Na, Si, P, and F ions released were significantly increased for all Maxxion R and Fuji IX groups (p < 0.0001). Cytotoxicity was increased only for Maxxion R with 5% and 10% of Biosilicate^®. A higher inhibition of S. mutans growth was observed for Maxxion R with 5% of Biosilicate^® (less than 100 CFU/mL), followed by Maxxion R with 10% of Biosilicate^® (p = 0.0053) and Maxxion R without the glass ceramic (p = 0.0093). Maxxion R and Fuji IX presented different behaviors regarding Biosilicate^® incorporation. The impacts on physico-mechanical and biological properties were different depending on the GIC, but therapeutic ion release was increased for both materials.

Background/Objectives: There are many commercial mouthrinses, used for a variety of purposes, including antiseptic activity. The objective of this study was to determine the antibacterial activity of various TheraBreath™ oral rinses against the cariogenic bacterium, Streptococcus mutans, and saliva-derived microbial communities, and their antibiofilm activity against S. mutans in vitro biofilms. Methods: Bactericidal activity against planktonic S. mutans was assessed by colony counting after 30 and 2 min exposures to mouthrinses. Ten saliva samples were exposed to mouthrinses for 30 s and plated aerobically on blood agar and Mitis Salivarius agar. Mature biofilms of S. mutans were treated with mouthrinses for 15 min followed by fluorescent vitality staining and polysaccharide measurement, followed by crystal violet staining for measurement of total biofilm remaining. Statistical analysis was performed using Kruskal–Wallis with Dunn’s multiple comparisons test comparing all mean ranks (α = 0.05). Results: TheraBreath™ Fresh Breath, Healthy Smile, and Dry Mouth exhibited no significant antibacterial activity. TheraBreath™ Healthy Gums showed antibacterial activity against S. mutans and microbes from saliva samples similar to Listerine^® Naturals at all exposure times. Whitening Fresh Breath showed intermediate killing of S. mutans after 30 min in liquid but not after 2 min or against salivary microbes. Live/Dead fluorescence vitality staining showed that Healthy Gums and Whitening Fresh Breath had antibacterial activity against mature biofilms of S. mutans statistically similar to Listerine^® Naturals and Colgate^® Total; however, Whitening Fresh Breath did not have significant killing compared to PBS. Conclusions: TheraBreath™ Healthy Gums demonstrated similar antiseptic activity levels to other antiseptic-claiming commercial rinses. Whitening Fresh Breath was comparable but unable to kill in short exposure times.