Study and evaluation of a gelatin- silver oxide nanoparticles releasing nitric oxide production of wound healing dressing for diabetic ulcer.

This study aimed to develop a novel Gelatin silver oxide material for releasing nitric oxide bionanocomposite wound dressing with enhanced mechanical, chemical, and antibacterial properties for the treatment of diabetic wounds. The gelatin- silver oxide nanoparticles (Ag2O-NP) bio nanocomposite was prepared using chitosan and gelatin polymers incorporated with silver oxide nanoparticles through the freeze-drying method. The samples were characterized using scanning electron microscopy (SEM) and X-ray diffraction (XRD) analysis. Results showed that the Ag2O-NP nanoparticles increased porosity, decreased pore size, and improved elastic modulus. The Ag2O-NP wound dressing exhibited the most effective antibacterial properties against Staphylococcus aureus and Escherichia coli. Among the samples, the wound dressing containing silver oxide nanoparticles demonstrated superior physical and mechanical properties, with 48% porosity, a tensile strength of 3.2 MPa, and an elastic modulus of 51.7 MPa. The fabricated wound dressings had a volume ratio of empty space to total volume ranging from 40% to 60%. In parallel, considering the complications of diabetes and its impact on the vascular system, another aspect of the research focused on developing a per2mediated wound dressing capable of releasing nitric oxide gas to regenerate damaged vessels and accelerate diabetic wound healing. Chitosan, a biocompatible and biodegradable polymer, was selected as the substrate for the wound dressing, and beta-glycerophosphate (GPß), tripolyphosphate (TPP), and per2mediated alginate (AL) were used as crosslinkers. The chitosan-alginate (CS-AL) wound dressing exhibited optimal characteristics in terms of hole count and uniformity in the scanning electron microscope test. It also demonstrated superior water absorption (3854%) and minimal air permeability. Furthermore, the CS-AL sample exhibited an 80% degradation rate after 14 days, indicating its suitability as a wound dressing. The wound dressing was loaded with S-nitrosoglutathione (GSNO) powder, and the successful release of nitric oxide gas was confirmed through the grease test, showing a peak at a wavelength of 540 nm. Subsequent investigations revealed that the treatment of human umbilical vein endothelial cells (HUVECs) with high glucose led to a decrease in the expression of PER2 and SIRT1, while the expression of PER2 increased, which may subsequently enhance the expression of SIRT1 and promote cell proliferation activity. However, upon treatment of the cells with the modified materials, an increase in the expression of PER2 and SIRT1 was observed, resulting in a partial restoration of cell proliferative activity. This comprehensive study successfully developed per2-mediated bio-nanocomposite wound dressings with improved physical, mechanical, chemical, and antibacterial properties. The incorporation of silver oxide nanoparticles enhanced the antimicrobial activity, while the released nitric oxide gas from the dressing demonstrated the ability to mitigate vascular endothelial cell damage induced by high glucose levels. These advancements show promising potential for facilitating the healing process of diabetic wounds by addressing complications associated with diabetes and enhancing overall wound healing.

Antibacterial and antibiofilm efficacy of Solanum lasiocarpum root extract synthesized silver/silver chloride nanoparticles against Staphylococcus haemolyticus associated with bovine mastitis.

Staphylococcus haemolyticus is a cause of bovine mastitis, leading to inflammation in the mammary gland. This bacterial infection adversely affects animal health, reducing milk quality and yield. Its emergence has been widely reported, representing a significant economic loss for dairy farms. Interestingly, S. haemolyticus exhibits higher levels of antimicrobial resistance than other coagulase-negative Staphylococci. In this study, we synthesized silver/silver chloride nanoparticles (Ag/AgCl-NPs) using Solanum lasiocarpum root extract and evaluated their antibacterial and antibiofilm activities against S. haemolyticus. The formation of the Ag/AgCl-NPs was confirmed using UV-visible spectroscopy, which revealed maximum absorption at 419 nm. X-ray diffraction (XRD) analysis demonstrated the crystalline nature of the Ag/AgCl-NPs, exhibiting a face-centered cubic lattice. Fourier transform infrared (FT-IR) spectroscopy elucidated the functional groups potentially involved in the Ag/AgCl-NPs synthesis. Transmission electron microscopy (TEM) analysis revealed that the average particle size of the Ag/AgCl-NPs was 10 nm. Antimicrobial activity results indicated that the minimum inhibitory concentration (MIC) and maximum bactericidal concentration (MBC) of the Ag/AgCl-NPs treatment were 7.82-15.63 µg/mL towards S. haemolyticus. Morphological changes in bacterial cells treated with the Ag/AgCl-NPs were observed under scanning electron microscopy (SEM). The Ag/AgCl-NPs reduced both the biomass of biofilm formation and preformed biofilm by approximately 20.24-94.66 % and 13.67-88.48 %. Bacterial viability within biofilm formation and preformed biofilm was reduced by approximately 21.56-77.54 % and 18.9-71.48 %, respectively. This study provides evidence of the potential of the synthesized Ag/AgCl-NPs as an antibacterial and antibiofilm agent against S. haemolyticus.

Biogenic synthesis, characterization, and in vitro biological investigation of silver oxide nanoparticles (AgONPs) using Rhynchosia capitata.

The current research aimed to study the green synthesis of silver oxide nanoparticles (AgONPs) using Rhynchosia capitata (RC) aqueous extract as a potent reducing and stabilizing agent. The obtained RC-AgONPs were characterized using UV, FT-IR, XRD, DLS, SEM, and EDX to investigate the morphology, size, and elemental composition. The size of the RC-AgONPs was found to be ~ 21.66 nm and an almost uniform distribution was executed by XRD analysis. In vitro studies were performed to reveal biological potential. The AgONPs exhibited efficient DPPH free radical scavenging potential (71.3%), reducing power (63.8 ± 1.77%), and total antioxidant capacity (88.5 ± 4.8%) to estimate their antioxidative power. Antibacterial and antifungal potentials were evaluated using the disc diffusion method against various bacterial and fungal strains, and the zones of inhibition (ZOI) were determined. A brine shrimp cytotoxicity assay was conducted to measure the cytotoxicity potential (LC50: 2.26 µg/mL). In addition, biocompatibility tests were performed to evaluate the biocompatible nature of RC-AgONPs using red blood cells, HEK, and VERO cell lines (< 200 µg/mL). An alpha-amylase inhibition assay was carried out with 67.6% inhibition. Moreover, In vitro, anticancer activity was performed against Hep-2 liver cancer cell lines, and an LC50 value of 45.94 µg/mL was achieved. Overall, the present study has demonstrated that the utilization of R. capitata extract for the biosynthesis of AgONPs offers a cost-effective, eco-friendly, and forthright alternative to traditional approaches for silver nanoparticle synthesis. The RC-AgONPs obtained exhibited significant bioactive properties, positioning them as promising candidates for diverse applications in the spheres of medicine and beyond.

Synthesis of novel silver-loaded clay AgF@Hec for the prevention of dental cariesin vitro.

Dental caries, a chronic infectious disease characterized by tooth mineral loss caused by plaque, is one of the major global public health problems. Silver diamine fluoride (SDF) has been proven to be a highly effective anti-caries drug due to its high bacterial inhibition and remineralization ability. However, the SDF solution is unstable, which immensely limits its clinical application. Therefore, new silver-load clay named AgF@Hec was designed by replacing the NH3with hectorite in this study. Fourier transform infrared spectroscopy and x-ray diffraction spectroscopy were employed to confirm the structure of AgF@Hec. Dynamic light scattering analysis was used to reveal the effect of different hectorite concentrations on the stability of AgF@Hec. Moreover, AgF@Hec exhibits significant remineralization and hardness recovery of the initial carious lesions. Bacteriostatic experiments also proved that it has a significant inhibitory effect onA. Viscosus, S. mutans, S. sanguinis, S. salivarius, Lactobacillus sp.and both gram-positive and gram-negative bacteria. We therefore believed that AgF@Hec should be a promising biomaterial that can be applied in the prevention of dental caries.

Exploiting the advantages of cationic copolymers and AgBr nanoparticles to optimize the antibacterial activity of chitosan.

Recently, the chitosan (CS)-based composites have attracted increasing attention for controlling and preventing the spread of pathogenic microorganisms. Herein, an amphiphilic copolymer containing epoxy and quaternary ammonium groups (PBGDBr) was synthesized via three common acrylate monomers. The epoxy groups of this copolymer were then crosslinked with the amino groups of CS to synthesize a natural/synthetic (PBGDBr-C) composite to increase the water solubility of CS under alkaline conditions and enhance its antibacterial activity based on chemical contact-type modes. Moreover, silver bromide nanoparticles (AgBr NPs)-decorated PBGDBr-C (AgBr@PBGDBr-C) composite was prepared, which aimed to endow the final AgBr@PBGDBr-C composite with a photodynamic antibacterial mode relying on the formation of Ag/AgBr nanostructures catalyzed by visible light on AgBr NPs. The results showed that the final composite possessed satisfactory bactericidal effects at concentrations higher than 64 and 128 µg/mL against Escherichia coli and Staphylococcus aureus, respectively. Additionally, The L929 cells treated with the final composite retained high cell viability (>80 %) at a concentration of 128 µg/mL, indicating its low toxicity to L929 cells. Overall, our synthetic strategy exploits a multi-modal system that enables chemical-photodynamic synergies to treat infections caused by pathogenic bacteria while delaying the development of bacterial resistance.

Comparison of the effects of fluoride varnish containing silver nanoparticles and conventional fluoride varnish on the surface microhardness of tooth enamel.

BACKGROUND: Nano-silver fluoride (NSF) has been introduced to improve enamel lesions. The effective use of varnishes is important in the prevention of dental caries. OBJECTIVES: The study aimed to compare the effect of conventional sodium fluoride varnish with the same varnish containing 1% and 2% silver nanoparticles (AgNP) on the surface microhardness of enamel. MATERIAL AND METHODS: The baseline surface microhardness of 40 premolar teeth was measured using a Vickers microhardness tester. After immersing the samples in a demineralizing agent for 24 h, the microhardness was measured again. In group B, a layer of conventional fluoride varnish was applied to the tooth surfaces using a microbrush with soft bristles, following the manufacturer's instructions. Groups C and D were treated with 1% and 2% NSF varnishes, respectively, while group A received no varnish. Surface microhardness tests were conducted on all specimens, including those previously tested. RESULTS: The microhardness of the enamel surface increased significantly in all 3 test groups compared to the microhardness after demineralization (p < 0.05). CONCLUSIONS: Conventional fluoride varnish and fluoride varnishes containing 1% and 2% AgNP are equally effective in remineralizing initial caries.

Silver vanadate nanomaterial incorporated into heat-cured resin and coating in printed resin - Antimicrobial activity in two multi-species biofilms and wettability.

OBJECTIVES: To incorporate the nanostructured silver vanadate decorated with silver nanoparticles (AgVO3) into denture base materials: heat-cured (HC) and 3D printed (3DP) resins, at concentrations of 2.5 %, 5 %, and 10 %; and to evaluate the antimicrobial activity in two multi-species biofilm: (1) Candida albicans, Candida glabrata, and Streptococcus mutans, (2) Candida albicans, Pseudomonas aeruginosa, and Staphylococcus aureus, and the wettability. METHODS: The AgVO3 was added to the HC powder, and printed samples were coated with 3DP with AgVO3 incorporated. After biofilm formation, the antimicrobial activity was evaluated by colony forming units per milliliter (CFU/mL), metabolic activity, and epifluorescence microscopy. Wettability was assessed by the contact angles with water and artificial saliva. RESULTS: In biofilm (1), HC-5 % and HC-10 % showed activity against S. mutans, HC-10 % against C. glabrata, and HC-10 % and 3DP-10 % had higher CFU/mL of C. albicans. 3DP-5 % had lower metabolic activity than the 3DP control. In biofilm (2), HC-10 % reduced S. aureus and P. aeruginosa, and HC-5 %, 3DP-2.5 %, and 3DP-5 % reduced S. aureus. 3DP incorporated with AgVO3, HC-5 %, and HC-10 % reduced biofilm (2) metabolic activity. 3DP-5 % and 3DP-10 % increased wettability with water and saliva. CONCLUSION: HC-10 % was effective against C. glabrata, S. mutans, P. aeruginosa, and S. aureus, and HC-5 % reduced S. mutans and S. aureus. For 3DP, 2.5 % and 5 % reduced S. aureus. The incorporation of AgVO3 into both resins reduced the metabolic activity of biofilms but had no effect on C. albicans. The wettability of the 3DP with water and saliva increased with the addition of AgVO3. CLINICAL SIGNIFICANCE: The incorporation of silver vanadate into the denture base materials provides antimicrobial efficacy and can prevent the aggravation of oral and systemic diseases. The incorporation of nanomaterials into printed resins is challenging and the coating is an alternative to obtain the inner denture base with antimicrobial effect.

Clinical evidence for silver diamine fluoride to reduce dentine hypersensitivity: A systematic review.

OBJECTIVE: To assess the clinical evidence for silver diamine fluoride (SDF) to reduce dentine hypersensitivity in adults. METHODS: Two independent researchers searched the English literature in five databases (Scopus, PubMed, Web of Science, EMBASE and the Cochrane Library) up to 15th July 2023 for clinical trials investigating the desensitising effect of professionally applied SDF to manage dentine hypersensitivity in adults aged 18 or above at any follow-up period. The primary outcome was the change in dentine hypersensitivity between baseline and follow-up visits after SDF application regarding any validated pain outcome measures. The Cochrane guidelines were used for the risk of bias assessment. RESULTS: Three hundred and thirty-one studies were identified, and four of them were finally included. Three of the included studies were rated as having a 'low risk' of bias. The SDF solution reduced dentine hypersensitivity in adults. The percentage reduction in dentine hypersensitivity ranged from 23 % to 56 % after a single application of SDF solution. Moreover, the SDF solution was more effective than potassium nitrate, potassium oxalate and glutaraldehyde plus hydroxyethyl methacrylate in reducing dentine hypersensitivity. Meta-analysis indicated a more significant reduction in visual analogue scales (1-10) by 1.35 (95 % CI:0.9-1.8; p<0.00001) after receiving the SDF application than controls. CONCLUSION: The included clinical trials showed that SDF solution reduced dentine hypersensitivity in adults. However, clinical trials are few, and their protocol varied from one another. Further well-designed clinical trials should be conducted to provide more evidence on its use to manage dentine hypersensitivity. CLINICAL SIGNIFICANCE: SDF is cleared as desensitizing agents by the US Food and Drug Administration to manage dentine hypersensitivity, which induces pain, limits food choice, and impacts the quality of life. Evidence from this systematic review informs clinicians and provides researchers insight for future research on SDF use for dentine hypersensitivity. THE INTERNATIONAL PROSPECTIVE REGISTER OF SYSTEMATIC REVIEWS (PROSPERO) REGISTRATION NUMBER: CRD42023462613.

Denture Adhesives Associated with Silver Vanadate: Antimicrobial Approach Against Multi- Species Biofilms on Acrylic Resin Surfaces.

Alternatives have been sought to add an antimicrobial property to denture adhesives. This study evaluated the antimicrobial potential of adhesives associated with nanostructured silver vanadate decorated with silver nanoparticles (ß-AgVO3). Specimens in acrylic resin were treated with the adhesives associated with ß-AgVO3 (1%, 2.5%, 5% and 10%). As control, specimens treated only with Ultra Corega Cream (UCC) or Ultra Corega Powder (UCP) adhesive were used. Multispecies biofilm of Candida albicans, Candida glabrata, Streptococcus mutans and Staphylococcus aureus was evaluated by counting colony forming units per milliliter (CFU/mL), colorimetric assay and fluorescence microscopy. The data were analyzed using the two-way analysis of variance (ANOVA) and Bonferroni multiple comparisons test (α=0.05). For both adhesives, a small amount of ß-AgVO3 (1%) completely inhibited S. mutans (P⟨0.05). For the other microorganisms, there was a reduction in metabolic activity and complete inhibition in the groups with intermediate or greater amounts of nanomaterial (P⟨0.05), except for C. albicans, which was reduced (P⟨0.05) but not completely inhibited in UCP. Microscopy that showed less biofilm in the groups with ß-AgVO3 and in the UCC than UCP. Denture adhesives in powder and cream form with ß-AgVO3 showed potential antimicrobial activity against multispecies biofilm. Powder adhesive showed higher biofilm formation.

Mechanical and microbiological properties of orthodontic resin modified with nanostructured silver vanadate decorated with silver nanoparticles (ßAgVO3).

OBJECTIVE: To investigate the impact of incorporating the antimicrobial nanomaterial ß-AgVO3 into orthodontic resin, focusing on degree of conversion, surface characteristics, microhardness, adhesion properties, and antimicrobial activity. METHODS: The 3 M Transbond XT resin underwent modification, resulting in three groups (Control, 2.5% addition, 5% addition) with 20 specimens each. Fourier transform infrared spectroscopy assessed monomer conversion. Laser confocal microscopy examined surface roughness, and microhardness was evaluated using Knoop protocols. Shear strength was measured before and after artificial aging on 36 premolar teeth. Microbiological analysis against S. mutans and S. sanguinis was conducted using the agar diffusion method. RESULTS: Degree of conversion remained unaffected by time (P = 0.797), concentration (P = 0.438), or their interaction (P = 0.187). The 5% group exhibited the lowest surface roughness, differing significantly from the control group (P = 0.045). Microhardness showed no significant differences between concentrations (P = 0.740). Shear strength was highest in the control group (P < 0.001). No significant differences were observed in the samples with or without thermocycling (P = 0.759). Microbial analysis revealed concentration-dependent variations, with the 5% group exhibiting the largest inhibition halo (P < 0.001). CONCLUSIONS: Incorporating ß-AgVO3 at 2.5% and 5% concentrations led to significant differences in surface roughness, adhesion, and antimicrobial activity. Overall, resin modification positively impacted degree of conversion, surface characteristics, microhardness, and antimicrobial activity. Further research is warranted to determine clinically optimal concentrations that maximize antimicrobial benefits while minimizing adverse effects on adhesion properties. CLINICAL SIGNIFICANCE: Incorporating ß-AgVO3 into orthodontic resin could improve patient quality of life by prolonging intervention durability and reducing the impact of cariogenic microorganisms. The study's findings also hold promise for the industry, paving the way for the development of new materials with antimicrobial properties for potential applications in the health sector.

The effect of silver nanoparticles in addition to sodium fluoride on remineralization of artificial root dentin caries.

BACKGROUND: Silver nanoparticle was developed to overcome the drawback of silver diamine fluoride. However, evidence is limited, especially in root caries. The aim of this study was to evaluate the remineralization effect of silver nanoparticles on root caries. MATERIALS AND METHODS: Fifty-five root human dentin slices size 5 × 5 mm2 from patients aged over 60 years old were immersed in demineralized solution to create artificial caries. Specimens were allocated into five groups according to the remineralizing agents: silver diamine fluoride (SDF), silver nanoparticles solution (AgNPs), silver nanoparticle solution followed by sodium fluoride varnish (AgNPs+NaF), sodium fluoride varnish (NaF), and tap water. After 8 days of pH-cycling challenge, the microhardness test, lesion depth evaluation, dentin surface morphology, and elemental analysis were performed. Data was analysed using F-test One-way ANOVA followed by Tukey's post hoc test and paired T-test. RESULTS: All test groups demonstrated a significantly higher microhardness value and lower lesion depth compared with the control group. AgNPs+NaF and NaF-treated groups showed lower efficacy than SDF. Crystal precipitation was presented in all groups composed of silver. CONCLUSION: Addition of fluoride varnish did not benefit for silver nanoparticles in preventing further demineralization. SDF provides the highest effectiveness in elderly root carious dentin.

Silver Compounds for Caries Management.

Silver metal and compounds have antibacterial properties, although their action's mechanisms are not fully understood. Scientists generally consider that silver disrupts the bacterial cell wall. It causes a structural change in the bacterial cell membrane and cytoplasm. It also stops deoxyribonucleic acid replication, resulting in inactivating enzymatic activity and cell death. The antimicrobial effect of silver-containing compounds relies on the release of bioactive silver ions. Hence, silver metal and compounds have been used in medicine to prevent infection for hundreds of years. Silver metal and compounds are also used as antibacterial agents in dentistry. Studies have shown that silver compounds are effective in the management of dental caries. Fluoride-containing silver compounds have been found in experiments to be beneficial at remineralising dental cavities. Silver diamine fluoride (SDF) can assist in preventing and arresting tooth cavities. The World Health Organization included SDF in its Model List of Essential Medicine for both adults and children in 2021. Clinicians also use SDF to manage dentine hypersensitivity as well as to inhibit growth of periodontal pathogens. However, traditional silver compounds cause tooth discolouration because of the silver-staining effect. These side effects of their applications depend on the amount applied and the frequency of application. Researchers are developing nanosilver fluoride and silver nanoparticles to overcome the staining. This review gives an overview of the antibacterial mechanism of silver compounds, namely silver nitrate, silver fluoride, SDF, silver nanoparticles, and nano silver fluoride for caries management. The outlook for the future development of silver compounds will be discussed.

Application of 20% silver nanoclusters in polymethacrylic acid on simulated dentin caries; its penetration depth and effect on surface hardness.

The aims of this study were: To evaluate the surface hardness of simulated dentin caries lesions treated with either silver nanoclusters (AgNCls) synthesized in polymethacrylic acid (PMAA) or 38% silver diammine fluoride (SDF), as well as observe the penetration of the treatment solutions into the simulated caries lesions. Dentin blocks 4 mm thick obtained from caries-free third molars were sectioned and then simulated caries lesions on the occlusal dentin surfaces were created. Each specimen (n = 8) was divided into four sections: (A) treated with 20% AgNCls/PMAA; (B) treated with SDF 38% (FAgamin, Tedequim, Cordoba, Argentina); (C) sound tooth protected by nail-varnish during artificial caries generation (positive control); and (D) artificial caries lesion without surface treatment (negative control). AgNCls/PMAA or SDF were applied on the simulated lesions with a microbrush for 10 s, then excess removed. The surface hardness was measured by means of Vickers indentation test. To trace the depth of penetration, up to 400 µm, of silver ions, elemental composition of the samples was observed using EDX, coupled with SEM, and measured every 50 µm from the surface towards the pulp chamber. Laser Induced Breakdown Spectroscopy (LIBS) was also employed to trace silver ion penetration; the atomic silver line 328.06 nm was used with a 60 µm laser spot size to a depth of 240 µm. Student's-t test identified significant differences between treatment groups for each depth and the Bonferroni test was used for statistical analysis of all groups (p < 0.05). Mean surface hardness values obtained were 111.2 MPa, 72.3 MPa, 103.3 MPa and 50.5 MPa for groups A, B, C and D respectively. There was a significant difference between groups A and C compared with groups B and D, the group treated with AgNCls/PMAA achieved the highest surface hardness, similar or higher than the sound dentin control. A constant presence of silver was observed throughout the depth of the sample for group A, while group B showed a peak concentration of silver at the surface with a significant drop beyond 50 µm. The 20% AgNCls/PMAA solution applied to simulated dentin caries lesions achieved the recovery of surface hardness equivalent to sound dentin with the penetration of silver ions throughout the depth of the lesion.

Effect of silver diamine fluoride upon the microbial community of carious lesions: A scoping review.

OBJECTIVES: To explore the effects of silver diamine fluoride (SDF) on the microbial community of carious lesions. DATA: Original studies evaluating the effect of SDF treatment on the microbial community of human carious lesions were included. SOURCES: A systematic search of English-language publications was performed in PubMed, EMBASE, Scopus, and Web of Science. Gray literature was searched in ClinicalTrials.gov and Google Scholar. STUDY SELECTION/RESULTS: This review included seven publications reporting the effects of SDF on microbial community of dental plaque or carious dentin, including the microbial biodiversity, relative abundance of microbial taxa, and predicted functional pathways of the microbial community. The studies on microbial community of dental plaque reported that SDF did not have a significant effect on both the within-community species diversity (alpha-diversity) and inter-community microbial compositional dissimilarity (beta-diversity) of the plaque microbial communities. However, SDF changed the relative abundance of 29 bacterial species of plaque community, inhibited carbohydrate transportation and interfered with the metabolic functions of the plaque microbial community. A study on the microbial community in dentin carious lesions reported that SDF affected its beta-diversity and changed the relative abundance of 14 bacterial species. CONCLUSION: SDF showed no significant effects on the biodiversity of the plaque microbial community but changed the beta-diversity of the carious dentin microbial community. SDF could change the relative abundance of certain bacterial species in the dental plaque and the carious dentin. SDF could also affect the predicted functional pathways of the microbial community. CLINICAL SIGNIFICANCE: This review provided comprehensive evidence on the potential effect of SDF treatment on the microbial community of carious lesions.

In situ detection of endogenous proteolytic activity and the effect of inhibitors on tooth root surface.

OBJECTIVES: The aim of this study was to clarify the distribution and activity of proteolytic enzymes and examine the inhibitory effects of various substances on this proteolytic activity on tooth root surfaces in situ. METHODS: Disk-shaped bovine tooth root samples were partly pretreated in acid solution (50 mM lactic acid buffer, pH 4.0) for 48 h. The fluorescence intensity of samples was detected with fluorescent substrate solution for collagenase and gelatinase using a stereoscopic fluorescence microscope for 60 min. The acid-pretreated and non-acid-pretreated root samples were treated with chlorhexidine (CHX), sodium fluoride (NaF), epigallocatechin gallate (EGCG), and calcium hydroxide (Ca(OH)2) for 10 min, and silver diamine fluoride (SDF) for 10, 30, and 60 s. These samples were immersed in the fluorescence substrate solution at pH 7.0, and the fluorescence intensity of samples was detected. The fluorescence intensity was calculated using analysis software. RESULTS: Gelatinase activity was detected in root samples. Gelatinase activity of the acid-pretreated side was significantly higher than that of the non-acid-pretreated side (1.63 times) at 60 min. CHX, EGCG, Ca(OH)2, and SDF decreased the gelatinase activity of root samples, while NaF had no effect. CONCLUSIONS: Gelatinase activity was detected from the root in situ and it was increased by acid-pretreatment. CHX, EGCG, Ca(OH)2, and SDF inhibited gelatinase activity. CLINICAL SIGNIFICANCE: Substances that inhibit proteolytic activity found in this research method will be useful for root caries prevention.

Time-dependent structural changes and hypermineralisation of artificially demineralised dentine following treatment with silver diammine fluoride and glass ionomer cement.

OBJECTIVES: This study aimed to evaluate the change of mineral content in dentine lesions over time and examine the effectiveness of the combined treatment with silver diammine fluoride (SDF) and glass ionomer cement (GIC). METHODS: Sixty bovine dentine specimens were divided into 4 groups: cont, Fuji, Safo, and Safo+Fuji. The specimens were imaged and measured using microcomputed tomography (microCT) at 7 time points: pre-demineralisation, after demineralisation for two weeks, immediately after treatment, 1 week, 2 weeks, 1 month, and 3 months after treatment. The 3-month group was evaluated with a light microscope, attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy, and scanning electron microscope (SEM)/energy-dispersive X-ray spectroscopy (EDS). Data were analysed by Dunn's test and Wilcoxon signed rank test with Bonferroni correction for microCT, and Kruskal-Wallis test and two-way analysis of variance for EDS characterisation. RESULTS: MicroCT images showed high mineral density beneath dentine lesions in Safo+Fuji. The mineral density at 600 µm in Safo+Fuji increased significantly over time, while Safo showed an opposite trend (adjusted p<0.005). In Safo+Fuji, EDS revealed significantly high energy of fluorine (p<0.05, at 300 µm) and a tendency towards high energy of calcium (p>0.05). However, Safo+Fuji showed lower energy of silver compared to Safo (p<0.001). ATR-FTIR revealed that phosphate groups had the highest peak at a depth between 300 and 400 µm in Safo+Fuji. CONCLUSIONS: Safo+Fuji was effective in remineralising the deep lesion in dentine after one and three months, and a hypermineralisation zone generated beneath the lesion demonstrated additional benefit in this study. CLINICAL SIGNIFICANCE: This long-term in vitro study showed that SDF+GIC treatment could strengthen the structure of decayed teeth when applied in the treatment of patients with advanced rampant caries.

Effect of silver nanoparticles associated with fluoride on the progression of root dentin caries in vitro.

OBJECTIVES: To assess the anti-proteolytic effect and potential to inhibit dentin root caries progression of a silver nanoparticle and fluoride solution (CNanoF) in comparison to silver diamine fluoride (SDF). METHODS: 48 specimens of root dentin artificial caries lesion were treated with 38% SDF, CNanoF, CNano or F (n = 6 per group). Ph cycling with demineralization and remineralization solutions simulated caries lesion progression. In addition, specimens were incubated with or without bacterial collagenase in the remineralization solution to induce dentin proteolytic degradation. Dentin degradation was assessed by weight loss rate and hydroxyproline (Hyp) release. Changes in cross-sectional microhardness, and lesion permeability and collagen integrity as determined by confocal laser scanning microscopy indicated potential for further demineralization inhibition. The effect of the solutions on the activity of metalloproteinases (MMP) -2 and -9 was also investigated. Statistical analysis consisted of ANOVA, Kruskal-Wallis, and linear mixed models with post-hoc pairwise Tukey, Dunn, and t-tests (α = 0.05). RESULTS: Treatment with SDF resulted in lower weight loss rate than did other solutions, but all groups showed similar Hyp release (p = 0.183). SDF resulted in greater microhardness at superficial layers of the caries lesions (p<0.05), while there were no differences among CNanoF, CNano, and F. Lesion permeability was similar among all groups after pH cycling (p>0.05), with or without the use of collagenase (p = 0.58). No statistically significant difference was noted among solutions regarding collagen integrity after pH cycling; however, SDF-treated dentin had a significant decrease in collagen integrity when collagenase was used (p = 0.003). Interestingly, only SDF was able to completely inactivate MMP-2 and -9. CONCLUSIONS: CNanoF and SDF both potentially prevent dentin degradation during caries lesion progression in vitro; however, SDF was more effective at inhibiting further tissue demineralization.

Antibacterial effect and impact on caries activity of nanosilver fluoride and silver diamine fluoride in dentin caries of primary teeth: a randomized controlled clinical trial.

BACKGROUND: The use of silver diamine fluoride (SDF) in caries treatment in children has increased despite the disadvantage of causing tooth discoloration. Nanosilver fluoride (NSF) is a possible alternative. This study aimed to assess the antibacterial effect of NSF and SDF and their impact on the activity of dentin caries in primary teeth. METHODS: Synthesis and characterization of the physical and biological properties of NSF were conducted. Fifty children aged 4-6 years with dentin caries (active caries corresponding to ICDAS code 5) in deciduous teeth were randomly assigned to treatment by NSF or SDF. Baseline assessment of Streptococcus mutans (S. mutans) and lactobacilli counts as CFU/mL in caries lesions was done, followed by the application of the agents. After one month, microbiological samples were recollected, and lesion activity was reassessed. Groups were compared using Mann-Whitney and Chi-Square tests, while intragroup comparisons were done using Wilcoxon and McNemar tests. Multilevel logistic regression analysis was used to assess the effect of different variables on the outcomes. RESULTS: There were 130 teeth in 50 children; mean ± SD age = 4.75 ± 0.76 years, 63% were posterior teeth. At the one-month follow-up appointment, both groups showed a significant decrease from baseline bacterial counts. There was a significant difference in the reduction of S. mutans between NSF and SDF (21.3% and 10.5%, respectively, p = 0.002), while not in lactobacilli (13.9% and 6.0%, respectively, p = 0.094). In both groups, there was a significant reduction in the number of active caries from baseline (p < 0.0001) with no significant difference between groups (percentage inactive = 64.4% and 63.4%, p = 0.903). Multilevel regression revealed non-significant differences in S. mutans and lactobacilli counts (AOR 1.281, p = 0.737 and 1.888, p = 0.341, respectively), and in the number of inactive lesions (AOR 1.355, p = 0.731) between groups. CONCLUSION: The short-term antibacterial efficacy of NSF was similar to that of SDF. In both groups there was a significant reduction of S. mutans and lactobacilli counts in active dentin caries, and two-thirds of the lesions became inactive with no differences between the two interventions. Further research is needed to investigate the long-term efficacy of NSF and its suitability for clinical use in caries management. TRIAL REGISTRATION: This trial was prospectively registered on the clinicaltrials.gov registry with ID: NCT05221749 on 03/02/2022.

Microbial silver resistance mechanisms: recent developments.

In this mini-review, after a brief introduction into the widespread antimicrobial use of silver ions and nanoparticles against bacteria, fungi and viruses, the toxicity of silver compounds and the molecular mechanisms of microbial silver resistance are discussed, including recent studies on bacteria and fungi. The similarities and differences between silver ions and silver nanoparticles as antimicrobial agents are also mentioned. Regarding bacterial ionic silver resistance, the roles of the sil operon, silver cation efflux proteins, and copper-silver efflux systems are explained. The importance of bacterially produced exopolysaccharides as a physiological (biofilm) defense mechanism against silver nanoparticles is also emphasized. Regarding fungal silver resistance, the roles of metallothioneins, copper-transporting P-type ATPases and cell wall are discussed. Recent evolutionary engineering (adaptive laboratory evolution) studies are also discussed which revealed that silver resistance can evolve rapidly in bacteria and fungi. The cross-resistance observed between silver resistance and resistance to other heavy metals and antibiotics in bacteria and fungi is also explained as a clinically and environmentally important issue. The use of silver against bacterial and fungal biofilm formation is also discussed. Finally, the antiviral effects of silver and the use of silver nanoparticles against SARS-CoV-2 and other viruses are mentioned. To conclude, silver compounds are becoming increasingly important as antimicrobial agents, and their widespread use necessitates detailed understanding of microbial silver response and resistance mechanisms, as well as the ecological effects of silver compounds. Figure created with BioRender.com.

Brucella species-induced brucellosis: Antimicrobial effects, potential resistance and toxicity of silver and gold nanosized particles.

Brucellosis is an endemic zoonotic disease caused by Brucella species, which are intramacrophage pathogens that make treating this disease challenging. The negative effects of the treatment regime have prompted the development of new antimicrobials against brucellosis. A new treatment modality for antibiotic-resistant microorganisms is the use of nanoparticles (NPs). In this study, we examined the antibacterial activities of silver and gold NPs (SNPs and GNPs, respectively), the resistance developed by Brucella melitensis (B. melitensis) and Brucella abortus (B. abortus) strains and the toxicity of both of these NPs in experimental rats. To test the bactericidal effects of the SNPs and GNPs, we used 22 multidrug-resistant Brucella isolates (10 B. melitensis and 12 B. abortus). The minimal inhibitory concentrations (MICs) of both types of NPs were determined utilizing the microdilution technique. To test the stability of resistance, 7 B. melitensis and 6 B. abortus isolates were passaged ten times in culture with subinhibitory concentrations of NPs and another ten times without NPs. Histopathological analysis was completed after rats were given 0.25, 0.5, 1, and 2 mg/kg NPs orally for 28 consecutive days. The MIC values (µg/ml) of the 10-nm SNPs and 20-nm GNPs against B. melitensis were 22.43 ± 2.32 and 13.56 ± 1.22, while these values were 18.77 ± 1.33 and 12.45 ± 1.59 for B. abortus, respectively. After extensive in vitro exposure, most strains showed no resistance to the 10-nm SNPs or 20-nm GNPs. The NPs and antibiotics did not cross-react in any of the evolved Brucella strains. SNPs and GNPs at doses below 2 mg/kg were not harmful to rat tissue according to organ histopathological examinations. However, a greater dose of NPs (2 mg/kg) harmed all of the tissues studied. The bactericidal properties of NPs are demonstrated in this work. Brucella strains develop similar resistance to SNPs and GNPs, and at low dosages, neither SNPs nor GNPs were hazardous to rats.