Low concentration zinc oxide nanoparticles enrichment enhances bacterial and pro-inflammatory resistance of calcium silicate-based cements.

Calcium silicate-based cement (CSC) is a commonly used material in endodontic treatment. However, it has limited antibacterial activity, especially for cases involving primary infections. Zinc oxide nanoparticles (ZnO-NPs) are recognized for their potential in biomedical applications due to their antibacterial properties and ability to reduce inflammation. This study aims to optimize CSC by incorporating ZnO-NPs to maintain its physical properties, enhance its antibacterial activity, and reduce the production of pro-inflammatory cytokines. ZnO-NPs were integrated into a commercial CSC (Endocem MTA) at 1 wt% (CSZ1) or 3 wt% (CSZ3). Setting time, compressive strength, and X-ray diffraction were then measured. In addition, pH, calcium ion release, and zinc ion release were measured for 7 days. Antibacterial activity against Enterococcus faecalis and viability of murine macrophages (RAW264.7) were determined using colorimetric assays. Gene expression levels of pro-inflammatory cytokines in lipopolysaccharide induced RAW264.7 were evaluated using quantitative polymerase chain reaction. Results were compared to an unmodified CSC group. In the CSZ3 group, there was a significant increase of approximately 12% in setting time and a reduction of about 36.4% in compressive strength compared to the control and CSZ1 groups. The presence of ZnO-NPs was detected in both CSZ1 and CSZ3. Both CSC and CSZ1 groups maintained an alkaline pH and released calcium ions, while zinc ions were significantly released in the CSZ1 group. Additionally, CSZ1 showed a 1.8-fold reduction of bacterial activity and exhibited around 85% reduction in colony-forming units compared to the CSC group. Furthermore, the CSZ1 group showed a more than 39% reduction in pro-inflammatory cytokine levels compared to the CSC group. Thus, enriching CSC with 1 wt% ZnO-NPs can enhance its antibacterial activity and reduce pro-inflammatory cytokines without showing any tangible adverse effects on its physical properties.

Effect of strontium substitution on functional activity of phosphate-based glass.

Phosphate-based glass (PBG) is a bioactive agent, composed of a glass network with phosphate as the primary component and can be substituted with various therapeutic ions for functional enhancement. Strontium (Sr) has been shown to stimulate osteogenic activity and inhibit pro-inflammatory responses. Despite this potential, there are limited studies that focus on the proportion of Sr substituted and its impact on the functional activity of resulting Sr-substituted PBG (PSr). In this study, focusing on the cellular biological response we synthesized and investigated the functional activity of PSr by characterizing its properties and comparing the effect of Sr substitution on cellular bioactivity. Moreover, we benchmarked the optimal composition against 45S5 bioactive glass (BG). Our results showed that PSr groups exhibited a glass structure and phosphate network like that of PBG. The release of Sr and P was most stable for PSr6, which showed favorable cell viability. Furthermore, PSr6 elicited excellent early osteogenic marker expression and inhibition of pro-inflammatory cytokine expression, which was significant compared to BG. In addition, compared to BG, PSr6 had markedly higher expression of osteopontin in immunocytochemistry, higher ALP expression in osteogenic media, and denser alizarin red staining in vitro. We also observed a comparable in vivo regenerative response in a 4-week rabbit calvaria defect model. Therefore, based on the results of this study, PSr6 could be identified as the functionally optimized composition with the potential to be applied as a valuable bioactive component of existing biomaterials used for bone regeneration.

Cerium oxide nanozymes confer a cytoprotective and bio-friendly surface micro-environment to methacrylate based oro-facial prostheses.

Poly-(methyl methacrylate) (PMMA) is the preferred biomaterial for orofacial prostheses used for the rehabilitation of naso-palatal defects. However, conventional PMMA has limitations determined by the complexity of the local microbiota and the friability of oral mucosa adjacent to these defects. Our purpose was to develop a new type of PMMA, i-PMMA, with good biocompatibility and better biological effects such as higher resistance to microbial adhesion of multiple species and enhanced antioxidant effect. The addition of cerium oxide nanoparticles to PMMA using a mesoporous nano-silica carrier and polybetaine conditioning, resulted in an increased release of cerium ions and enzyme mimetic activity, without tangible loss of mechanical properties. Ex vivo experiments confirmed these observations. In stressed human gingival fibroblasts, i-PMMA reduced the levels of reactive oxygen species and increased the expression of homeostasis-related proteins (PPARg, ATG5, LCI/III). Furthermore, i-PMMA increased the levels of expression of superoxide dismutase and mitogen-activated protein kinases (ERK and Akt), and cellular migration. Lastly, we demonstrated the biosafety of i-PMMA using two in vivo models: skin sensitization assay and oral mucosa irritation test, respectively. Therefore, i-PMMA offers a cytoprotective interface that prevents microbial adhesion and attenuates oxidative stress, thus supporting physiological recovery of the oral mucosa.

Changes in mechanical and bacterial properties of denture base resin following nanoceria incorporation with and without SBA-15 carriers.

Poly (methyl methacrylate) (PMMA) is a commonly used material for the fabrication of biomedical appliances. Although PMMA has several advantages, it is susceptible to microbial insults with practical use. Therefore, different bioactive nanomaterials, such as nanoceria (CeN), have been proposed to enhance the properties of PMMA. In this study, we investigated the effect of the incorporation of CeN into PMMA with and without the use of mesoporous silica nanoparticle (SBA-15) carriers. The unmodified PMMA specimens (control, CTRL) were compared to groups containing SBA-15, CeN, and the synthesized SBA-15 impregnated with CeN (SBA-15@CeN) at different loading percentages. The mechanical and physical properties of the different SBA-15@CeN groups and their effects on cell viability were investigated, and the optimal CeN concentration was identified accordingly. Our results revealed that flexural strength was significantly (P < 0.01) reduced in the SBA-15@CeN3× group (containing 3-fold the CeN wt. %). Although the surface microhardness increased with the increase in the wt. % of SBA-15@CeN, cell viability was significantly reduced (P < 0.001). The SBA-15@CeN1× group had the optimal concentration and displayed significant resistance to single-and multispecies microbial colonization. Finally, the enzymatic activity of CeN was significantly high in the SBA-15@CeN1× group. The proinflammatory markers (IL-6, IL-1ß, TNF-α, CD80, and CD86) showed a significant (P < 0.001) multifold reduction in lipopolysaccharide-induced RAW cells treated with a 5-day eluate of the SBA-15@CeN1× group. These results indicate that the addition of SBA-15@CeN at 1.5 wt % improves the biological response of PMMA without compromising its mechanical properties.

Novel Osteogenic and Easily Handled Endodontic Calcium Silicate Cement Using Pluronic F127 Hydrogel.

Calcium silicate cement (CSC) is widely used as an endodontic material in clinical applications such as direct pulp capping, pulpotomy, or root canal. CSC has good biocompatibility, sealing properties, and the ability to enhance hard tissue regeneration. However, the disadvantage of CSC is the difficulty in handling when placing it into endodontic tissue due to the long setting time. Several attempts have been made to improve handling of CSC; however, these methods were limited by osteogenic properties. To overcome such a disadvantage, this study investigated the use of Pluronic F127 (F127) for the development easy-to-handle novel endodontic CSCs with osteogenic properties. In this case, different concentrations of F127 (5%, 10%, 20%, 30%, and 40%) were implemented to generate CSC specimens H5, H10, H20, H30, and H40, respectively. Calcium ion was continuously released for 28 days. In addition, each group resulted in apatite formation for 28 days corresponding to calcium ion release. The concentration of F127 showed opposite relationships with water solubility and compressive strength. The H20 group showed a high level of osteogenic activity compared to other groups at 14 days. Mineralization of the H20 group was higher than that of the other groups. This study indicates that the novel F127-based hydrogel with CSC can potentially be used as endodontic filler.

Improvement of the mechanical and biological properties of bioactive glasses by the addition of zirconium oxide (ZrO2 ) as a synthetic bone graft substitute.

In this study, mechanical properties of bioactive glass (BAG) synthetic bone graft substitute was improved by containing ZrO2 (ZrO2 -BAG), while maintaining advantageous biological properties of BAG such as osteoinductive and osteoconductive properties. The ZrO2 -BAG was produced by adding ZrO2 in the following proportions to replace Na2 O in 45S5 BAG: 1% (Zr1-BAG), 3% (Zr3-BAG), 6% (Zr6-BAG), and 12% (Zr12-BAG). Properties including XRD, XPS, SEM, DSC, fracture toughness, and Vickers microhardness were evaluated. To assess the biological properties, Ca/P apatite formation, ion release, degradation rate, cell proliferation, ALP activity (ALP), and alizarin red S staining assay (ARS) were evaluated. Also, expression of osteogenic differentiation markers, Osteopontin (OPN), confirmed by immunofluorescence staining. Finally, an in vivo test was carried out to by implanting ZrO2 -BAG into the subcutaneous tissue of rats. The results of each test were statistically analyzed with one-way ANOVA followed by Tukey's post hoc statistical test. Amorphous ZrO2 -BAG was successfully produced with increased mechanical properties as the ZrO2 content was increased. Additionally, ZrO2 -BAG exhibited a slower ion release and degradation rate compare to BAG without ZrO2 . Bioactivity of ZrO2 -BAG was confirmed with apatite layer formed on the surface, significantly higher proliferation rate and significantly enhanced ALP and the degree of ARS of the cells compare to respective controls. The tissue reactions observed in the in vivo study showed neo-formed vessels after implantation of ZrO2 -BAG.

Development of a Bioactive Flowable Resin Composite Containing a Zinc-Doped Phosphate-Based Glass.

Flowable resins used for dental restoration are subject to biofilm formation. Zinc has antibacterial properties. Thus, we prepared a zinc-doped phosphate-based glass (Zn-PBG) to dope a flowable resin and evaluated the antibacterial activity of the composite against Streptococcus mutans (S. mutans) to extrapolate the preventative effect toward secondary caries. The composites were prepared having 0 (control), 1.9, 3.8, and 5.4 wt.% Zn-PBG. The flexural strength, elastic modulus, microhardness, depth of cure, ion release, inhibition zone size, and number of colony-forming units were evaluated and analyzed using ANOVA. The flexural strength of the control was significantly higher than those of Zn-PBG samples (p < 0.05). However, all samples meet the International Standard, ISO 4049. The microhardness was not significantly different for the control group and 1.9 and 3.8 wt.% groups, but the 5.4 wt.% Zn-PBG group had a significantly lower microhardness (p < 0.05). Further, the composite resins increasingly released P, Ca, Na, and Zn ions with an increase in Zn-PBG content (p < 0.05). The colony-forming unit count revealed a significant reduction in S. mutans viability (p < 0.05) with increase in Zn-PBG content. Therefore, the addition of Zn-PBG to flowable composite resins enhances antibacterial activity and could aid the prevention of secondary caries.

Osteogenic Properties of Novel Methylsulfonylmethane-Coated Hydroxyapatite Scaffold.

Despite numerous advantages of using porous hydroxyapatite (HAp) scaffolds in bone regeneration, the material is limited in terms of osteoinduction. In this study, the porous scaffold made from nanosized HAp was coated with different concentrations of osteoinductive aqueous methylsulfonylmethane (MSM) solution (2.5, 5, 10, and 20%) and the corresponding MH scaffolds were referred to as MH2.5, MH5, MH10, and MH20, respectively. The results showed that all MH scaffolds resulted in burst release of MSM for up to 7 d. Cellular experiments were conducted using MC3T3-E1 preosteoblast cells, which showed no significant difference between the MH2.5 scaffold and the control with respect to the rate of cell proliferation (p > 0.05). There was no significant difference between each group at day 4 for alkaline phosphatase (ALP) activity, though the MH2.5 group showed higher level of activity than other groups at day 10. Calcium deposition, using alizarin red staining, showed that cell mineralization was significantly higher in the MH2.5 scaffold than that in the HAp scaffold (p < 0.0001). This study indicated that the MH2.5 scaffold has potential for both osteoinduction and osteoconduction in bone regeneration.

Effect of wet storage on the bioactivity of ultraviolet light- and non-thermal atmospheric pressure plasma-treated titanium and zirconia implant surfaces.

The aim of this study was to evaluate whether combining two treatments to avoid biological aging of the surface of titanium and zirconia implants; i.e., storage in an aqueous solution after ultraviolet light (UV) or non-thermal atmospheric pressure plasma (NTP) treatment, yielded surface bioactivity comparable to that following post-15-min UV or NTP treatment storage under air or immediately after UV or NTP treatment. Grade IV titanium discs modified by large grit sand-blasting and acid-etching (SLA) and smooth zirconia discs were irradiated with UV or NTP and their surface properties were evaluated immediately and after storage for 8 weeks in distilled H2O (dH2O) and a sealed container under air. Approximately 15-30 nm-sized nano-protrusions were formed only on SLA surfaces in dH2O immediately after UV or NTP treatment. Immediate dH2O storage after UV or NTP treatment prevented hydrocarbon contamination and maintained elevated amounts of Ti and Zr. After 8 weeks, unlike zirconia, protein adsorption, cellular adhesion, and cytoskeletal development of MC3T3-E1 cells on SLA surfaces stored in dH2O immediately after UV treatment were further exceeding those immediately after UV or NTP treatments. UV treatment of SLA implants followed by wet storage can not only maintain but also strengthen bioactivity during shelf storage.

Bioactive resin-based composite with surface pre-reacted glass-ionomer filler and zwitterionic material to prevent the formation of multi-species biofilm.

OBJECTIVE: This study evaluated the synergetic effect between surface pre-reacted glass-ionomer (SPRG) filler and 2-methacryloyloxyethyl phosphorylcholine (MPC), for inhibiting multi-species biofilm formation, while maintaining or even improving the original beneficial features of SPRG-filled resin-based composite (RBC). METHODS: MPC (1.5-10wt%) was incorporated into commercial SPRG-filled RBC. Then, the inherent properties of RBC, and ion release and acid-neutralising properties associated with SPRG were investigated. Further, protein adsorptions and bacterial adhesion and viability on the SPRG-filled RBC surfaces were studied using four kinds of oral bacteria; Streptococcus mutans, Actinomyces naeslundii, Veillonella parvula, and Porphyromonas gingivalis. Finally, the thickness and biomass of the human saliva-derived biofilm model cultured on test and control samples were analysed. RESULTS: Addition of MPC content resulted in decreased flexural strength and wettability of SPRG-filled RBC. SPRG-filled RBC released significantly higher amounts of multiple ions as contents of MPC increased. Meanwhile, SPRG-filled RBC with 5-wt% MPC significantly improved acid-neutralising properties than those of other test and control samples (P<0.001). SPRG-filled RBC with 3wt% MPC significantly reduced the amount of adsorbed bovine serum albumin and proteins from the brain heart infusion medium as compared to the control (P<0.01). A similar trend was observed in the attachment of four types of bacteria and multi-species biofilm (P<0.01). SIGNIFICANCE: Despite limitation in terms of deteriorations of some physical properties, addition of 3% MPC to SPRG-filled RBC leads to inhibition of the attachment of multi-species bacteria on its surface, as well as inhibition of biofilm growth. Moreover, the original important bioactive features of SPRG-filled RBC such as ion release and acid neutralisations are either maintained or improved upon adding MPC.

Comparing Properties of Variable Pore-Sized 3D-Printed PLA Membrane with Conventional PLA Membrane for Guided Bone/Tissue Regeneration.

The aim of this study was to fabricate bioresorbable polylactide (PLA) membranes by 3D printing and compare their properties to those of the membranes fabricated by the conventional method and compare the effect of different pore sizes on the properties of the 3D-printed membranes. PLA membranes with three different pore sizes (large pore-479 µm, small pore-273 µm, and no pore) were 3D printed, and membranes fabricated using the conventional solvent casting method were used as the control group. Scanning electron microscopy (SEM) and micro-computed tomography (µ-CT) were taken to observe the morphology and obtain the porosity of the four groups. A tensile test was performed to compare the tensile strength, elastic modulus, and elongation at break of the membranes. Preosteoblast cells were cultured on the membranes for 1, 3 and 7 days, followed by a WST assay and SEM, to examine the cell proliferation on different groups. As a result, the 3D-printed membranes showed superior mechanical properties to those of the solvent cast membranes, and the 3D-printed membranes exhibited different advantageous mechanical properties depending on the different pore sizes. The various fabrication methods and pore sizes did not have significantly different effects on cell growth. It is proven that 3D printing is a promising method for the fabrication of customized barrier membranes used in GBR/GTR.

Novel anti-biofouling light-curable fluoride varnish containing 2-methacryloyloxyethyl phosphorylcholine to prevent enamel demineralization.

We evaluated the efficacy of light-curable fluoride varnish (LCFV) that contains 2-methacryloyloxyethyl phosphorylcholine (MPC) in terms of anti-biofouling properties and prevention of tooth enamel demineralization. MPC was mixed with and incorporated into LCFV at 0 (control), 1.5, 3.0, 5.0, 10.0, 20.0, and 40.0 weight percentage (wt%). Addition of high wt% of MPC resulted in increased film thickness and decreased the degree of conversion, indicating loss of the advantageous properties of LCFV. Addition of 1.5, 3, or 5 wt% MPC significantly reduced the amount of bovine serum albumin adsorbed from a solution and proteins adsorbed from brain heart infusion medium compared to the control (P < 0.001). A similar pattern was observed for bacterial adhesion: significantly less Streptococcus mutans cells adhered on the surface of LCFV with 1.5, 3, or 5 wt% MPC (P < 0.001) than on the control, and similar results were obtained for Actinomyces naeslundii and Streptococcus sanguinis adherence to LCFV with 3 wt% MPC. Finally, bacterial adhesion, surface microhardness loss, and the depth of demineralization were substantially lower on bovine tooth enamel surface coated with LCFV containing 3 wt% of MPC than in the control treatment (0 wt% MPC). Therefore, this novel LCFV containing a low concentration of MPC (e.g., 3 wt%) would be effective in anti-biofouling while maintaining the important advantageous features of light-curable fluoride in preventing demineralization.

Novel anti-biofouling bioactive calcium silicate-based cement containing 2-methacryloyloxyethyl phosphorylcholine.

Calcium silicate-based cements (CSCs) are commonly used for endodontic procedures; however, their antibacterial effects are limited. The objective of this study was to develop a 2-methacryloyloxyethyl phosphorylcholine (MPC)-incorporated CSC with improved antibacterial properties, while maintaining the original advantageous features of CSC. MPC was incorporated into a commercial CSC (Endocem MTA) at 0 wt% (control), 1.5%, 3.0 wt%, 5.0 wt%, 7.5 wt%, and 10 wt%. The setting time, compressive strength, water sorption, and glycerol contact angle were measured. Protein absorption was measured and bacterial adhesion on the surface was evaluated using Enterococcus faecalis. The bactericidal effect was examined by the disc diffusion test. Mineralization ability was assessed based on calcium ion deposition, as assessed by alizarin red staining, after immersion into Hank's balanced salt solution for 7 days. High concentrations of MPC in CSC (7.5 wt% and 10 wt%) increased the setting time, reduced compressive strength, and reduced wettability. MPC (3 wt%) had greater protein repellent and anti-biofouling effects than those of control and test materials (P < 0.001). However, no bactericidal effect was observed for any control or test materials. There was greater calcium ion deposition on the surface of MPC-supplemented CSC than on the control (P < 0.001). The addition of 3 wt% MPC polymer to CSC confers protein-repellent properties and reduced bacterial attachment, with the potential for improved mineralization.

Effects of thermoforming on the physical and mechanical properties of thermoplastic materials for transparent orthodontic aligners.

OBJECTIVE: The aim of this systematic multiscale analysis was to evaluate the effects of thermoforming on the physical and mechanical properties of thermoplastic materials used to fabricate transparent orthodontic aligners (TOAs). METHODS: Specimens were fabricated using four types of thermoplastic materials with different thicknesses under a thermal vacuum. Transparency, water absorption and solubility, surface hardness, and the results of three-point bending and tensile tests were evaluated before and after thermoforming. Data were analyzed using one-way analysis of variance and Student's t-test. RESULTS: After thermoforming, the transparency of Duran and Essix A+ decreased, while the water absorption ability of all materials; the water solubility of Duran, Essix A+, and Essix ACE; and the surface hardness of Duran and Essix A+ increased. The flexure modulus for the 0.5-mm-thick Duran, Essix A+, and eCligner specimens increased, whereas that for the 0.75-/1.0-mm-thick Duran and eClginer specimens decreased. In addition, the elastic modulus increased for the 0.5-mm-thick Essix A+ specimens and decreased for the 0.75-mm-thick Duran and Essix ACE and the 1.0-mm-thick Essix ACE specimens. CONCLUSIONS: Our findings suggest that the physical and mechanical properties of thermoplastic materials used for the fabrication of TOAs should be evaluated after thermoforming in order to characterize their properties for clinical application.