Antibacterial activities of phytochemicals against Porphyromonas gingivalis with and without experimental fluoride varnish for periodontal disease prevention.

We aimed to evaluate the antibacterial activity of phytochemicals with or without an experimental fluoride varnish against Porphyromonas gingivalis. Five phytochemicals, chrysophanol (CHR), emodin (EMO), anthrarufin (ANT), bavachalcone (BCC), and isobavachromene (IBC), were tested using agar diffusion, minimal inhibition concentration (MIC), and minimum bacterial concentration (MBC) assays. We also assessed the cell viability and cytotoxicity of phytochemicals. All phytochemicals showed clear inhibition zones in the agar diffusion test. The inhibition zones of all phytochemical-containing fluoride varnishes were similar to or larger than that of the positive control, excluding that of 1 mM EMO. With or without the fluoride varnish, BCC exhibited the lowest MIC and MBC levels. Cell viability was high in the presence of all phytochemicals except 200 µM EMO. In conclusion, BCC was most effective as a phytochemical alone, while all phytochemical-containing fluoride varnishes inhibited P. gingivalis growth without cytotoxicity.

Effect of adhesive components in experimental fluoride varnish on fluoride release within 30 days in vitro study.

We aimed to determine whether adhesive components could increase the release time of effective fluoride concentration from an experimental fluoride varnish applied to bovine teeth. An experimental fluoride varnish containing 5% sodium fluoride (EX1) was prepared and combined with 35% hydroxyethyl methacrylate (HEMA) (EX2), 5% glutaraldehyde (EX3), or 35% HEMA/5% glutaraldehyde mixture (EX4). Two commercially available fluoride varnishes were used for comparison. Each group was applied to bovine incisors, and the fluoride release and pH were monitored for 30 days. Cell viability analysis, scanning electron microscopy, and energy-dispersive spectroscopy were performed. EX4 released the highest and most effective concentration of fluoride for the longest period and reached neutral pH at the earliest; the release was maintained for up to 30 days without cytotoxicity. In conclusion, EX4 is considered to be the most effective varnish to prevent dental caries.

Effectiveness of a collagen matrix seal and xenograft in alveolar ridge preservation: an experimental study in dogs.

Majority of previous studies on alveolar ridge preservation (ARP) used collagen membranes as barrier membranes, and further evidence for ARP in dehiscent extraction sockets with a deproteinized bovine bone mineral (DBBM) and matrix is needed. The aim of this study is to assess the impact of non-cross linked collagen membranes (membrane) and crosslinked collagen matrices (matrix) on ARP using DBBM in extraction sockets with buccal dehiscence. In six mongrel dogs, the mesial roots of three mandibular premolars (P2, P3, and P4) were extracted 1 month after dehiscence defect induction. Two experimental groups were randomly assigned: (1) DBBM with a membrane (DBBM/membrane group) and (2) DBBM with a matrix (DBBM/matrix group). Three-dimensional (3D) volumetric, microcomputed tomography (µCT), and histologic analyses were performed to assess the ridge preservation. Both groups were effective to maintain the ridge width (p > 0.05), and the DBBM/matrix group showed more favorable soft tissue regeneration and bone quality in the histological analysis (p = 0.05). Based on these results, DBBM/matrix could be better choice for ARP in cases of buccal dehiscence defects.

Antibacterial Activity and Sustained Effectiveness of Calcium Silicate-Based Cement as a Root-End Filling Material against Enterococcus faecalis.

Several calcium silicate cement (CSC) types with improved handling properties have been developed lately for root-end filling applications. While sealing ability is important, a high biocompatibility and antimicrobial effects are critical. This study aimed to conduct a comparative evaluation of the antimicrobial efficacy and sustained antibacterial effectiveness against Enterococcus faecalis (E. faecalis) of commercially available CSCs mixed with distilled water (DW) and chlorhexidine (CHX). Various products, viz., ProRoot mixed with DW (PRW) or with CHX (PRC), Endocem mixed with DW (EW) or with CHX (EC), and Endocem premixed (EP) syringe type, were used. While antibacterial activity against E. faecalis was evaluated using a direct contact method, the specimens were stored in a shaking incubator for 30 d for antibacterial sustainability. The cytotoxicity was evaluated using a cell counting kit-8 assay in human periodontal ligament stem cells. The antibacterial activities of EP, EW, and EC were greater than those of PRC and PRW (p < 0.05). The antibacterial sustainability of EP was the highest without cytotoxicity for up to 30 days (p < 0.05). In conclusion, the pre-mixed injectable type EP was most effective in terms of antibacterial activity and sustained antibacterial effectiveness without cytotoxicity.

Evaluation of alveolar ridge preservation in sockets with buccal dehiscence defects using two types of xenogeneic biomaterials: An in vivo experimental study.

OBJECTIVES: Alveolar ridge preservation (ARP) has been extensively investigated in various preclinical and clinical studies, yielding favorable results. We aim to evaluate the effects of ARP using collagenated bovine bone mineral (CBBM) alone or particulated bovine bone mineral with a non-cross-linked collagen membrane (PBBM/NCLM) in tooth extraction sockets with buccal dehiscence in an experimental dog model. MATERIALS AND METHODS: The mesial roots of three mandibular premolars (P2, P3, and P4) were extracted from six mongrel dogs 4 weeks after inducing dehiscence defects. ARP was randomly performed using two different protocols: 1) CBBM alone and 2) PBBM/NCLM. Three-dimensional (3D) volumetric, micro-computed tomography, and histological analyses were employed to determine changes over a span of 20 weeks. RESULTS: In 3D volumetric and radiographic analyses, CBBM alone demonstrated similar effectiveness to PBBM/NCLM in ARP (p > .05). However, in the PBBM/NCLM group (3.05 ± 0.60 mm), the horizontal ridge width was well maintained 3 mm below the alveolar crest compared with the CBBM group (2.11 ± 1.01 mm, p = .002). CONCLUSION: Although the radiographic changes in the quality and quantity of bone were not significant between the two groups, the use of PBBM/NCLM resulted in greater horizontal dimensions and more favorable maintenance of the ridge profile.

Gingival biotype modification with collagen matrix or autogenous subepithelial connective tissue graft: Histologic and volumetric analyses in a beagle model.

Objectives: To evaluate the volumetric effect and biocompatibility of porcine tendon-derived type I collagen matrix graft (CG) in gingival biotype modification (GBM) compared with subepithelial connective tissue graft (SCTG) in a beagle model. Methods: Surface analysis using scanning electron microscopy and a collagen degradation assay of CG was performed in vitro. Six adult dogs were used in in vivo experiment, and each received autologous SCTG or CG at the anterior side. Histometric and three-dimensional digital volume analyses were conducted to compare quantitative changes in CG and SCTG in GBM. Immunohistochemical analysis was performed for the qualitative evaluation of CG compared to SCTG. Results: CG had a double-layered structure, and its degradation was slower than that of other well-reported materials. No critical problems were associated with the healing procedure. Changes in gingival thickness and volume in the CG and SCTG groups were equivalent, with no significant differences between the groups. Type I collagen and vascular endothelial growth factor expression levels were similar in both groups. Significance: CG and SCTG had equivalent potential for GBM in terms of quantity and quality. Additionally, CG could be used as a reasonable substitute for SCTG, making surgery convenient and predicting successful clinical outcomes.

Enhanced Bone Formation by Rapidly Formed Bony Wall over the Bone Defect Using Dual Growth Factors.

BACKGROUND: In guided bone regeneration (GBR), there are various problems that occur in the bone defect after the wound healing period. This study aimed to investigate the enhancement of the osteogenic ability of the dual scaffold complex and identify the appropriate concentration of growth factors (GF) for new bone formation based on the novel GBR concept that is applying rapid bone forming GFs to the membrane outside of the bone defect. METHODS: Four bone defects with a diameter of 8 mm were formed in the calvaria of New Zealand white rabbits each to perform GBR. Collagen membrane and biphasic calcium phosphate (BCP) were applied to the bone defects with the four different concetration of BMP-2 or FGF-2. After 2, 4, and 8 weeks of healing, histological, histomorphometric, and immunohistochemical analyses were conducted. RESULTS: In the histological analysis, continuous forms of new bones were observed in the upper part of bone defect in the experimental groups, whereas no continuous forms were observed in the control group. In the histomorphometry, The group to which BMP-2 0.5 mg/ml and FGF-2 1.0 mg/ml was applied showed statistically significantly higher new bone formation. Also, the new bone formation according to the healing period was statistically significantly higher at 8 weeks than at 2, 4 weeks. CONCLUSION: The novel GBR method in which BMP-2, newly proposed in this study, is applied to the membrane is effective for bone regeneration. In addition, the dual scaffold complex is quantitatively and qualitatively advantageous for bone regeneration and bone maintenance over time.

Photobiomodulation-Based Synergic Effects of Pt-Coated TiO2 Nanotubes and 850 nm Near-Infrared Irradiation on the Osseointegration Enhancement: In Vitro and In Vivo Evaluation.

Photobiomodulation (PBM) therapy is known to have the potential to improve bone regeneration after implant surgery. However, the combinatory effect of the nanotextured implant and PBM therapy on osseointegration has not yet been proved. This study evaluated the photobiomodulation-based synergistic effects of Pt-coated titania nanotubes (Pt-TiO2 NT) and 850 nm near-infrared (NIR) light on osteogenic performance in vitro and in vivo. The FE-SEM and the diffuse UV-Vis-NIR spectrophotometer were used to perform the surface characterization. The live-dead, MTT, ALP, and AR assays were tested to perform in vitro tests. The removal torque testing, the 3D-micro CT, and the histological analysis were used to conduct in vivo tests. The live-dead and MTT assay resulted in Pt-TiO2 NTs being biocompatible. The ALP activity and AR assays demonstrated that the combination of Pt-TiO2 NT and NIR irradiation significantly enhanced osteogenic functionality (p < 0.05). The results of in vivo test, employing the removal torque testing, the 3D-micro CT, and histological analysis, showed overall improved outcomes; however, no significant difference was observed between the control and experimental groups (p > 0.05). Therefore, we confirmed the possibility of the combination of Pt-TiO2 NT and NIR light as a promising technology for implant surgery in dentistry.

Improving Bone Formation by Guided Bone Regeneration Using a Collagen Membrane with rhBMP-2: A Novel Concept.

We examined whether recombinant human bone morphogenetic protein-2 (rhBMP-2) when applied to collagen membranes, would reinforce them during guided bone regeneration. Four critical cranial bone defects were created and treated in 30 New Zealand white rabbits, including a control group, critical defect only; group 1, collagen membrane only; group 2, biphasic calcium phosphate (BCP) only; group 3, collagen membrane + BCP; group 4, collagen membrane with rhBMP-2 (1.0 mg/mL); group 5, collagen membrane with rhBMP-2 (0.5 mg/mL); group 6, collagen membrane with rhBMP-2 (1.0 mg/mL) + BCP; and group 7, collagen membrane with rhBMP-2 (0.5 mg/mL) + BCP. After a 2-, 4-, or 8-week healing period, the animals were sacrificed. The combination of collagen membranes with rhBMP-2 and BCP yielded significantly higher bone formation rates compared to the other groups (control group and groups 1-5 < groups 6 and 7; p < 0.05). A 2-week healing period yielded significantly lower bone formation than that at 4 and 8 weeks (2 < 4 = 8 weeks; p < 0.05). This study proposes a novel GBR concept in which rhBMP-2 is applied to collagen membranes outside instead of inside the grafted area, thereby inducing quantitatively and qualitatively enhanced bone regeneration in critical bone defects.

Evaluation of the machinability and machining accuracy of polymer-based CAD/CAM blocks using merlon fracture test model.

The aims of this study were to estimate the machinability and machining accuracy of polymer-based CAM blocks using the merlon fracture test model specified in ISO 18675: 2022. Three hybrid disc blanks (MazicDuro, HC Disk, and Enamic) and three polymethyl methacrylate (PMMA) disc blanks (PMMA Disk, PMMA Block, and MazicTemp Hybrid) were tested in this study. The machinability was evaluated by assigning scores according to the fracture range of merlons with areas divided by ten vertical planes. The machining accuracy was evaluated by superimposing methods via CAD reference data and CAD specimen data. Within the limits of this study, a thickness of 0.3 mm is recommended for the clinical application of polymer-based CAD/CAM blocks in dental restorations that require superior machinability and accuracy. In addition, the machinability and machining accuracy tests of polymer-based CAM blocks are expected to provide guidelines for preparing accurate dental restorations.

A Comparative Study of Additive and Subtractive Manufacturing Techniques for a Zirconia Dental Product: An Analysis of the Manufacturing Accuracy and the Bond Strength of Porcelain to Zirconia.

This study was aimed at preparing zirconia samples via additive manufacturing (AM) and subtractive manufacturing (SM) and testing the following aspects: (1) the manufacturing accuracy of the zirconia samples and (2) the bond strength of porcelain to zirconia to evaluate the applicability of the zirconia fabricated by AM in dental clinics. We used three milling machines for SM (AR, K5, and UP) and a 3D printer for AM (AO). The manufacturing accuracy of the zirconia specimen in the internal and marginal areas was evaluated by superimposing techniques to calculate the root mean square (RMS) values. The bond strengths of porcelain to zirconia prepared via SM and AM were measured using a universal testing machine. The internal and marginal RMS values of the zirconia prepared by AM (AO) were within the range of those of the zirconia prepared by SM (AR, K5, and UP). Moreover, the bond strength value of the zirconia prepared by AM (35.12 ± 4.09 MPa) was significantly higher than that of the zirconia prepared by SM (30.26 ± 5.20 MPa). Therefore, AM technology has significant potential for applications in dentistry.

Oral Microbiome Using Colocasia antiquorum var. esculenta Extract Varnish in a Mouse Model with Oral Gavage of P. gingivalis ATCC 53978.

Background and Objective: There is increasing interest in preventing periodontitis using natural products. The purpose of this study was to investigate the effect of Colocasia antiquorum var. esculenta (CA) varnish on the oral microbiome and alveolar bone loss in a mouse periodontitis model. Materials and Methods: Antibacterial activity against Porphyromonas gingivalis (P. gingivalis) ATCC 53978 and cell cytotoxicity using CCK-8 on L929 cells were measured. Balb/c mice were assigned into five groups (negative control, positive control, CA in drinking water, varnish, and CA varnish). P. gingivalis was administered to the mice by oral gavage three times. After sacrifice, the oral microbiome and the levels of the inflammatory cytokine IL-1ß and matrix metalloproteinase-9 were analyzed. Alveolar bone loss was measured using micro-computed tomography. Results: CA extract showed an antibacterial effect against P. gingivalis (p < 0.05) and showed no cytotoxicity at that concentration (p > 0.05). Although alpha diversity of the oral microbiome did not statistically differ between the groups (p > 0.05), the relative abundance of dominant bacteria tended to be different between the groups. The inflammatory cytokine IL-1ß was reduced in the CA varnish group (p < 0.05), and no difference was observed in MMP-9 expression and alveolar bone loss (p > 0.05). Conclusions: CA varnish did not affect the overall microflora and exhibited an anti-inflammatory effect, suggesting that it is possibility a suitable candidate for improving periodontitis.

Shear bond strengths of various resin cements between three types of adherends and bovine teeth with and without thermocycling.

This study evaluated the shear bond strengths of various types of resin cements between three types of adherends (composite resin, metal, and ceramic) and bovine teeth with and without thermocycling. A conventional resin cement (Variolink N), two adhesive resin cements (PANAVIA F 2.0, Multilink N), and three self-adhesive resin cements (MAXCEM ELITE, Rely X Unicem 2, Speed CEM) were used. The adherends were cemented on the superficial dentin of bovine incisors using each resin cement. Herein, 10 specimens from each group were thermocycled 5,000 times, and the other 10 were stored without thermocycling. With the resin and ceramic adherends, the shear bond strengths of Rely X Unicem 2 were significantly higher than those of the other resin cements both with and without thermocycling (p<0.05). With the metal adherend, the shear bond strengths were not significantly different among the cement groups, except MAXCEM ELITE, which showed the lowest strength.

Visible Light-Mediated Sustainable Antibacterial Activity and Osteogenic Functionality of Au and Pt Multi-Coated TiO2 Nanotubes.

The visible light reactions of noble metal-based photocatalysts have been increasingly utilized to investigate their antibacterial activities. Furthermore, the photoreactions at various visible light wavelengths for specific combinations of titania nanotubes and noble metal nanoparticles have been found to promote osteogenic functionality. In this investigation, a novel multi-coating combination of noble metals (gold and platinum) on titania nanotubes was assessed using plasmonic photocatalysis and low-level laser therapy at 470 and 600 nm. The results showed that this coating on the nanotubes promoted antibacterial activity and osteogenic functionality. The order in which the gold and platinum coatings were layered onto the titania nanotubes strongly affected the osteogenic performance of the human mesenchymal stem cells. These results have identified a new approach for the development of efficient novel combinations of noble metal nanoparticles and titania nanotubes with visible light responses, sustainable antimicrobial activity, and osteogenic functionality.

Effects of Colocasia antiquorum var. Esculenta Extract In Vitro and In Vivo against Periodontal Disease.

Background and Objectives: Periodontal disease is a chronic inflammatory disease in which gradual destruction of tissues around teeth is caused by plaque formed by pathogenic bacteria. The purpose of this study was to evaluate the potential of 75% ethanol extract of Colocasia antiquorum var. esculenta (CA) as a prophylactic and improvement agent for periodontal disease in vitro and in vivo. Materials and Methods: The antimicrobial efficacy of CA against Porphyromonas gingivalis (P. gingivalis, ATCC 33277) was evaluated using minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) test, and cytotoxicity was confirmed by CCK-8 assay. For the in vivo study, P. gingivalis was applied by oral gavage to BALB/c mice. Forty-two days after the first inoculation of P. gingivalis, intraoral swabs were taken for microbiome analysis, and the mice were sacrificed to evaluate the alveolar bone loss. Results: The MIC of CA against P. gingivalis was 31.3 µg/mL, the MBC was 62.5 µg/mL, with no cytotoxicity. The diversity of the oral microbiome decreased in the positive control group, while those of the VA (varnish) and VCA (varnish added with CA) groups increased as much as in the negative control group, although the alveolar bone loss was not induced in the mouse model. Conclusions: CA showed antibacterial effects in vitro, and the VA and VCA groups exhibited increased diversity in the oral microbiome, suggesting that CA has potential for improving periodontal disease.

Inhibitory Effect of Asplenium incisum on Bacterial Growth, Inflammation, and Osteoclastogenesis.

Background and Objectives:Asplenium incisum, a natural plant, is known to possess numerous pharmacological and biochemical properties. However, the inhibitory effect of A. incisum against Porphyromonas gingivalis and other factors related to periodontal disease have not yet been demonstrated. This study aimed to investigate the potential of A. incisum extract as a phytotherapeutic candidate for improving periodontal diseases by assessing its antibacterial, anti-inflammatory, and anti-osteoclastogenic activities. Materials and Methods: The inhibition of proliferation of P. gingivalis by A. incisum and the sustainability of its antibacterial activity were evaluated in this study. The production of inflammatory cytokines (tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)) and nitric oxide (NO) from lipopolysaccharide-stimulated RAW 264.7 cells was assessed using an enzyme-linked immunosorbent assay. To identify the anti-osteoclastogenic activity, tartrate-resistant acid phosphatase (TRAP) staining and TRAP activity analyses were performed on bone marrow macrophages. Results: The proliferation of P. gingivalis was significantly inhibited by A. incisum (p < 0.001), and the antibacterial activity was sustained for up to 3 days. A. incisum showed anti-inflammatory activities by significantly decreasing the release of TNF-α, IL-6 (p < 0.05), and NO (p < 0.01). In addition, A. incisum significantly suppressed TRAP-positive cells and TRAP activity (at 30 µg/mL, p < 0.01) without causing any cytotoxicity (p > 0.05). Conclusions:A. incisum showed antibacterial, anti-inflammatory, and anti-osteoclastogenic activities, suggesting it has strong therapeutic potential against periodontal diseases.

In Vivo Study for Clinical Application of Dental Stem Cell Therapy Incorporated with Dental Titanium Implants.

The aim of this study was to investigate the behavior of dental-derived human mesenchymal stem cells (d-hMSCs) in response to differently surface-treated implants and to evaluate the effect of d-hMSCs on local osteogenesis around an implant in vivo. d-hMSCs derived from alveolar bone were established and cultured on machined, sandblasted and acid-etched (SLA)-treated titanium discs with and without osteogenic induction medium. Their morphological and osteogenic potential was assessed by scanning electron microscopy (SEM) and real-time polymerase chain reaction (RT-PCR) via mixing of 5 × 106 of d-hMSCs with 1 mL of Metrigel and 20 µL of gel-cell mixture, which was dispensed into the defect followed by the placement of customized mini-implants (machined, SLA-treated implants) in New Zealand white rabbits. Following healing periods of 2 weeks and 12 weeks, the obtained samples in each group were analyzed radiographically, histomorphometrically and immunohistochemically. The quantitative change in osteogenic differentiation of d-hMSCs was identified according to the type of surface treatment. Radiographic analysis revealed that an increase in new bone formation was statistically significant in the d-hMSCs group. Histomorphometric analysis was in accordance with radiographic analysis, showing the significantly increased new bone formation in the d-hMSCs group regardless of time of sacrifice. Human nuclei A was identified near the area where d-hMSCs were implanted but the level of expression was found to be decreased as time passed. Within the limitations of the present study, in this animal model, the transplantation of d-hMSCs enhanced the new bone formation around an implant and the survival and function of the stem cells was experimentally proven up to 12 weeks post-sacrifice.

Antibacterial, anti-inflammatory, and anti-osteoclastogenic activities of Colocasia antiquorum var. esculenta: Potential applications in preventing and treating periodontal diseases.

This study aimed to investigate the inhibitory effects of Colocasia antiquorum var. esculenta (CA) on Porphyromonas gingivalis (P. gingivalis) growth, inflammation, and osteoclastogenesis. CA was effective in inhibiting the growth of P. gingivalis when applied together with an experimental fluoride varnish. CA also significantly decreased the release of interleukin-6, tumor necrosis factor-α, and nitric oxide from lipopolysaccharide-induced RAW 264.7 cells. No significant differences in viability were noted between the cells treated with CA and the controls. In addition, CA significantly attenuated osteoclast differentiation on bone marrow macrophages. In conclusion, CA inhibited the growth of P. gingivalis and showed anti-inflammatory and anti-osteoclastogenic effects. Therefore, CA may have the potential to act as a novel natural agent for preventing periodontitis.

Visible Light-Enhanced Antibacterial and Osteogenic Functionality of Au and Pt Nanoparticles Deposited on TiO2 Nanotubes.

This study aimed at evaluating the visible light mediated antimicrobial and osteogenic applications of noble metal, such as gold (Au) and platinum (Pt) coated titania (TiO2) nanotubes (NTs). In this study, the Au and Pt nanoparticles (NPs) were deposited on anodized 100 nm TiO2 NTs by ion plasma sputtering. The Au and Pt NPs were mainly deposited on the top surface layer of TiO2 NTs and showed light absorbance peaks around the 470 and 600 nm visible light region used in this study, as seen from the surface characterization. From the results of antibacterial activity test, Au and Pt NPs that were deposited on TiO2 NTs showed excellent antibacterial activity under 470 nm visible light irradiation due to the plasmonic photocatalysis based on the localized surface plasmon resonance effect of the Au and Pt NPs. In addition, alkaline phosphate activity test and quantitative real-time PCR assay of osteogenic related genes resulted that these NPs promoted the osteogenic functionality of human mesenchymal stem cells (hMSCs) under 600 nm visible light irradiation, because of the synergic effect of the photothermal scattering of noble metal nanoparticles and visible light low-level laser therapy (LLLT). Therefore, the combination of noble metal coated TiO2 NTs and visible light irradiation would be expected to perform permanent antibacterial activity without the need of an antibacterial agent besides promoting osteogenic functionality.

Inhibitory effects on Streptococcus mutans of antibacterial agents mixed with experimental fluoride varnish.

We investigated the inhibitory effect of antibacterial agents mixed with experimental fluoride varnish (EFV) on Streptococcus mutans. The antibacterial agents used were (1 and 10) mM of xanthorrhizol, bakuchiol, bavachalcone, isobavachromene, and bavachromene. Agar diffusion tests were performed on S. mutans (1.1×1010 CFU/mL), using antibacterial agents without and with EFV. Bavachalcone showed the highest inhibition zone without and with EFV at both (1 and 10) mM (p<0.05). All EFV with antibacterial agents showed greater inhibition and semi-inhibition zones than EFV alone (p<0.05). The cell viability of each antibacterial agent was not significantly different from the vehicle controls (p>0.05), except xanthorrhizol and bakuchiol at 1 mM. All antibacterial agents were effective, while antibacterial agents with EFV co-formulations were more effective than EFV alone. Bavachalcone was the most effective agent against S. mutans, indicating its potential usefulness with fluoride varnish in preventing dental caries.