Sonic-assisted antibacterial photodynamic therapy: a strategy for enhancing lateral canal disinfection.

BACKGROUND: Bacterial infections in lateral canals pose challenges for root canal treatment. This in vitro study aims to evaluate the antibacterial efficacy of sonic-assisted methylene blue mediated antimicrobial photodynamic therapy (MB-aPDT) against Enterococcus faecalis (E. faecalis) in infected lateral canals. METHODS: Sixty-five premolars infected with E. faecalis in lateral canals were randomly divided into five groups (n = 13) and treated with : (1) 5.25% NaOCl (positive control); (2) Saline (negative control); (3) Sonic-assisted MB-aPDT; (4) 3% NaOCl + MB-aPDT; (5) 3% NaOCl + sonic-assisted MB-aPDT, respectively. The antibacterial efficacy was evaluated by the colony- counting method (CCM) and scanning electronic microscope (SEM). RESULTS: Both 5.25% NaOCl and the 3% NaOCl + sonic-assisted MB-aPDT exhibited the most effective while comparable antibacterial effects without significant statistical difference (P > 0.05). Furthermore, the antibacterial effect of the 3% NaOCl + MB-aPDT group was significantly higher compared to that of the sonic-assisted MB-aPDT group (P < 0.05). The SEM results demonstrated notable morphological alterations in E. faecalis across all experimental groups, except for the negative control group. CONCLUSION: The concentration of NaOCl can be reduced to a safe level while preserving its antibacterial efficacy through the synergism with the sonic-assisted MB-aPDT in this study.

Antimicrobial Photodynamic Effect of Cross-Kingdom Microorganisms with Toluidine Blue O and Potassium Iodide.

Streptococcus mutans (S. mutans) and Candida albicans (C. albicans) are prominent microbes associated with rapid and aggressive caries. In the present study, we investigated the antimicrobial efficacy, cytotoxicity, and mechanism of toluidine blue O (TBO)-mediated antimicrobial photodynamic therapy (aPDT) and potassium iodide (KI). The dependence of KI concentration, TBO concentration and light dose on the antimicrobial effect of aPDT plus KI was determined. The cytotoxicity of TBO-mediated aPDT plus KI was analyzed by cell counting kit-8 (CCK-8) assay. A singlet oxygen (1O2) probe test, time-resolved 1O2 detection, and a 1O2 quencher experiment were performed to evaluate the role of 1O2 during aPDT plus KI. The generation of iodine and hydrogen peroxide (H2O2) were analyzed by an iodine starch test and Amplex red assay. The anti-biofilm effect of TBO-mediated aPDT plus KI was also evaluated by counting forming unit (CFU) assay. KI could potentiate TBO-mediated aPDT against S. mutans and C. albicans in planktonic and biofilm states, which was safe for human dental pulp cells. 1O2 measurement showed that KI could quench 1O2 signals, implicating that 1O2 may act as a principal mediator to oxidize excess iodide ions to form iodine and H2O2. KI could highly potentiate TBO-mediated aPDT in eradicating S. mutans and C. albicans due to the synergistic effect of molecular iodine and H2O2.

Laser activated irrigation with SWEEPS modality reduces concentration of sodium hypochlorite in root canal irrigation.

INTRODUCTION: Shockwave enhanced emission photoacoustic streaming (SWEEPS) of Er: YAG laser has been proposed to improve the disinfecting efficacy of endodontic treatments. The purpose of this in vitro study was to compare the effects of different concentrations of sodium hypochlorite (NaOCl) combined with SWEEPS root canal irrigation on the removal of Enterococcus faecalis (E.faecalis) in infected bovine root canals. MATERIALS AND METHODS: Forty-eight bovine root canals infected with E.faecalis were randomly divided into six groups. The root canals were irrigated by normal saline (NS) in combination with conventional needle irrigation (CI) , NS and different concentrations of NaOCl (0.5%, 1.0%, 2.0%, and 5.25%) in combination with SWEEPS. Then, the remaining live microbes from all specimens before and after root canal irrigation were collected and recovered via plate counting of the colony-forming unit (CFU). The CFU data were statistically analyzed by the Kruskal-Wallis H test. RESULTS: After root canal irrigation, the viable count in each group was significantly reduced (P < 0.05). The bacterial reduction in NS+CI and NS+SWEEPS groups was significantly lower than that in other groups (P < 0.05), and there was no significant difference among the other groups, or between these two groups (P > 0.05). CONCLUSIONS: This study showed that SWEEPS used for root canal irrigation can enhance the effect of low-concentration NaOCl while ensuring the antimicrobial effect.

Hydrogen peroxide potentiates antimicrobial photodynamic therapy in eliminating Candida albicans and Streptococcus mutans dual-species biofilm from denture base.

BACKGROUND: Candida albicans (C.albicans) is the primary pathogen of denture biofilm. Moreover, it could establish a cross-kingdom relationship with bacteria to enhance its virulence and resistance to antifungal drugs. This study aimed to investigate the efficacy of antimicrobial photodynamic therapy (aPDT) in combination with hydrogen peroxide (H2O2) against C.albicans and Streptococcus mutans (S.mutans) dual-species biofilm formed on polymethyl methacrylate (PMMA) disk, and explore its involved mechanisms. METHODS: C.albicans and S.mutans were grown on PMMA disk for 48 h to form biofilm and received different treatments. The treatments included:1) phosphate-buffered saline (PBS) group,2) 100 mM H2O2 group,3) aPDT group,4) aPDT+ H2O2 and 5) H2O2+aPDT group. Colony forming units (CFU), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and scanning electron microscope (SEM) were used to evaluate the antimicrobial effects. Extracellular polysaccharide substance (EPS) production and observation, cell permeability of biofilm, and uptake of toluidine blue O (TBO) by biofilm were assessed to investigate the involved mechanism. RESULTS: There was no significant difference between PBS group and H2O2 group in viable microorganisms and metabolic activity of biofilm. The treatment protocols containing aPDT group reduced microorganism numbers and metabolic activity when compared to PBS group or H2O2 group (P<0.05). H2O2+aPDT treatment showed the highest antimicrobial efficacy in comparison with other treatments (P<0.05). Pretreatment with H2O2 could decrease EPS production and enhance cell permeability, leading to increased TBO uptake in biofilm. CONCLUSION: Pretreatment with H2O2 improved aPDT efficiency in eliminating dual-species biofilm from PMMA disk by reducing EPS amount, enhancing cell permeability, and increasing TBO uptake.

Pretreatment of Root Canal with Photodynamic Therapy Facilitates Adhesion, Viability and Differentiation of Stem Cells of the Apical Papilla.

This study was to test the hypothesis that root canal pretreated with photodynamic therapy (PDT) would promote stem cells from the apical papilla (SCAP) adhesion, proliferation and differentiation without affecting smear layer removal and microhardness of root canal. Standardized root canals were randomized into four groups (n = 30/group): (1) sodium hypochlorite(NaOCl) group, (2) NaOCl + ethylene diaminetetraacetic acid (EDTA) group, (3) NaOCl + PDT group, (4) NaOCl + EDTA + PDT group. After treatments, smear layer removal and microhardness of root canal were evaluated. SCAP with hydroxyapatite-based scaffolds were seeded into root canals for 7 days. SCAP adhesion was observed by scanning electron microscope (SEM), and viable cells were calculated by CellTiter-Glo Luminescent kit. Platelet-derived growth factor (PDGF) and vascular endothelial growth factor (VEGF) expression of SCAP were evaluated by Quantitative Reverse Transcriptase-Polymerase Chain Reaction. There was no significant difference in the smear layer removal and microhardness of root dentin between the groups with and without PDT treatment (P > 0.05). SCAP with elongated cytoplasmic processes and cell-cell contact were observed on the dentin surfaces treated with PDT. Elevated cell viability, PDGF and VEGF expression were found in root canal treated with PDT (P < 0.05). Under the experimental conditions, PDT could provide positive microenvironment for SCAP growth.

Disinfection of Cariogenic Pathogens in Planktonic Lifestyle, Biofilm and Carious Dentine with Antimicrobial Photodynamic Therapy.

Antimicrobial photodynamic therapy (aPDT) has been recommended for clinical application. Its antibacterial effect on bacteria remained in dentinal tubule was seldom investigated. Here, we evaluated the antibacterial effects of aPDT on Streptococcus mutans (S. mutans) and Lactobacillus acidophilus (L. acidophilus) in planktonic lifestyle, biofilm and carious dentine. Mono-species biofilms or dentinal caries formed on human dentine slices or slabs. Bacterial suspension, biofilms and dentine caries were treated with 0.1 mg mL-1 toluidine Blue O followed by irradiation with a light emission diode (λ - 635 ± 10 nm; 500 mW; 31.5 J cm-2 ; 60 s) and 0.12% chlorhexidine (CHX), respectively. Residual bacteria were determined by microbial culture analysis and scanning electron microscopy (SEM). One-way analysis of variance (ANOVA) was performed to detect the significance of the variables. Both treatments significantly reduced the number of L. acidophilus in planktonic state, biofilm and carious dentine (P < 0.05). For S. mutans, CHX was only bactericidal against suspension (P < 0.05), while aPDT was effective on both suspension and biofilm (P < 0.05) while not for dentin caries (P > 0.05). Under the experimental conditions assessed, aPDT could be an alternative disinfection method for superficial layer of caries cavity. Its disinfection on bacteria in dentinal tubule of deep layer was deficient.

Antimicrobial effects of photodynamic therapy with antiseptics on Staphylococcus aureus biofilm on titanium surface.

BACKGROUND: S. aureus biofilm plays a predominant role in the establishment and development of peri-implantitis. It is suggested to combine different modalities as peri-implantitis treatment. The aim of this study was to evaluate the disinfection efficacy of combined application of antiseptics with PDT on Staphylococcus aureus (S. aureus) biofilm formed on titanium(Ti) disks with different surface roughness. METHODS: S. aureus biofilm was incubated on polished and sandblasted large-grit acid-etched (SLA) Ti surfaces for 48 h. 72 contaminated Ti disks (36 polished, 36 SLA) were randomly divided into 6 different groups as follows: (a) PBS, (b) 0.2% chlorhexidine digluconate (CHX), (c) 3% hydrogen peroxide (H2O2), (d) PDT, (e) 0.2% CHX + PDT, and (f) 3% H2O2+PDT. Colony forming unit (CFU) was measured to determine antimicrobial effects. Biofilm structure was assessed using scanning electron microscopy (SEM) and confocal laser scanning microscope (CLSM). RESULTS: All disinfection methods significantly reduced bacteria amounts compared to control group on both polished and SLA Ti surfaces (P < 0.001). PDT demonstrated stronger decontamination ability in eliminating S. aureus from Ti surfaces than CHX and H2O2 did (P < 0.05). The combined CHX or H2O2 with PDT treatment were more effective in bacterial disinfection than a single administration of these treatments (P < 0.001). CONCLUSION: The combination of CHX or H2O2 administration with PDT was more effective in eradicating S. aureus on both polished and SLA Ti disks in comparison with either treatment alone, suggesting that combined usage of antiseptics with PDT could be a more efficient method for the treatment of peri-implantitis.

Disinfect Porphyromonas gingivalis Biofilm on Titanium Surface with Combined Application of Chlorhexidine and Antimicrobial Photodynamic Therapy.

Various antimicrobial modalities have been proposed to treat peri-implantitis but resulted in limited outcomes. The aim of this in vitro study was to evaluate the disinfection efficacy of combined application of chlorhexidine digluconate (CHX) and antimicrobial photodynamic therapy (aPDT) of titanium surfaces previously contaminated with Porphyromonas gingivalis biofilm. P. gingivalis biofilms were grown on 32 polished and 32 sandblasted large-grit acid-etched (SLA) titanium surfaces. Titanium disks were allocated into four groups as follows: (1) immersed in phosphate-buffered saline (PBS), (2) immersed in 0.2% CHX, (3) application of aPDT and (4) immersed in 0.2% CHX and subsequent aPDT. Residual bacteria were determined by microbial culture analysis and by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM) imaging. Combination protocol (CHX+ aPDT) was the most effective in eradicating P. gingivalis (P < 0.05) on both polished and SLA surfaces. There was no significant difference in the number of remaining P. gingivalis between polished titanium disks and the SLA ones in four groups (P > 0.05). Under the limitation of this study, combined technique of preceding application of CHX and subsequent aPDT was shown to be an efficient method in reducing P. gingivalis numbers in both polished and SLA titanium surfaces.

The efficacy of platelet-rich fibrin as a scaffold in regenerative endodontic treatment: a retrospective controlled cohort study.

BACKGROUND: Blood Clot (BC) or platelet concentrates have been used as scaffold in regenerative endodontic treatment (RET). The aim of this retrospective study was to compare the performance of platelet-rich fibrin (PRF) with BC in inducing root development and periapical lesion healing after tooth revascularization. METHODS: Five patients receiving RET using PRF as a scaffold were matched 1:1 to a previous cohort of 5 patients who underwent tooth revascularization by provoking periapical bleeding. Clinical signs and symptoms were examined at follow-ups. Periapical lesion healing and root development were monitored radiographically. The resolution of clinical signs and symptoms as well as periapical radiolucency was observed in all patients (100%). RESULTS: Root elongation, dentinal wall thickening and apex closure were found in most cases (80% in both groups). There was no significant difference between the groups in terms of clinical sign resolution, root development and periapical healing. CONCLUSIONS: Within the limits of this study, PRF achieved comparable outcomes to BC in terms of clinical sign and symptom resolution, periapical lesion healing and continued root development in RET.

Histologic and Immunohistochemical Findings of a Human Immature Permanent Tooth with Apical Periodontitis after Regenerative Endodontic Treatment.

Specimens of human immature permanent teeth after regenerative endodontic treatment (RET) are sparse. This case report describes the histologic and immunohistochemical findings of tissue formed in the canal space of a human immature permanent tooth with apical periodontitis after RET. A patient presenting with immature human permanent tooth #29 with apical periodontitis underwent RET. At the 10-month follow-up visit, radiographic examination revealed complete resolution of the periapical lesion, marked narrowing of the apical foramen, increased thickness of the canal walls, and minimal lengthening of the root. Notably, the tooth regained pulp sensibility. Tooth #29 was extracted for orthodontic reasons and processed for histologic and immunohistochemical examination. The canal space was filled with newly formed cementumlike tissue, bonelike tissue, and fibrous connective tissue. The apical closure, thickness, and length increment of the root were caused by the deposition of cementumlike tissue without dentin. Furthermore, neurons and nerve fibers were observed in the canal space; this observation was confirmed by immunohistochemistry. Based on the findings in the present case, after RET, the newly formed tissues in the canal space of the human immature permanent tooth with apical periodontitis were primarily fibrous connective tissue, cementumlike tissue, and bonelike tissue. Nerve regeneration was identified.

[Effect of the self-etching adhesives system on human pulp fibroblast].

OBJECTIVE: To compare and evaluate the biocompatibility of three kinds of dentin bonding agents Xeno III (XO), Adper Prompt (AP), Single bond2 (SB) through cell culture in vitro. METHODS: Three kinds of dentin bonding agents (XO, AP, SB) were applied on the surface of the dental slices which were 5.0 mm in diameter and 0.5 mm in depth. By immersing the slices into the DMEM culture medium, the maceration extracts were obtained. Normal dental pulps of teenagers were collected and human pulp fibroblast was cultured using tissue explant method. The fifth generation pulp cells were exposed to culture medium containing different concentrations of maceration extracts (100.0%, 50.0%, 25.0%, 12.5%) for 24, 72, 120 h. At last, MTT method was used to evaluate the cytotoxicity of the dentin bonding agents on human pulp fibroblast. RESULTS: The results showed that all three kinds of dentin bonding systems had cytotoxicity to human pulp fibroblast in different degree in vitro. The cytotoxicity of XO and AP was less than SB. The difference was statistically significant (P<0.05). CONCLUSION: The results of cell culture in vitro indicated that total-etching adhesives system has more irritation to pulp than self-etching adhesives system.