Periodontal ligament stem cell-derived exosome-loaded Emodin mediated antimicrobial photodynamic therapy against cariogenic bacteria.

BACKGROUND: This study was conducted to investigate the efficiency of periodontal ligament (PDL) stem cell-derived exosome-loaded Emodin (Emo@PDL-Exo) in antimicrobial photodynamic therapy (aPDT) on Streptococcus mutans and Lactobacillus acidophilus as the cariogenic bacteria. MATERIALS AND METHODS: After isolating and characterizing PDL-Exo, the study proceeded to prepare and verify the presence of Emo@PDL-Exo. The antimicrobial effect, anti-biofilm activity, and anti-metabolic potency of Emo, PDL-Exo, and Emo@PDL-Exo were then evaluated with and without irradiation of blue laser at a wavelength of 405 ± 10 nm with an output intensity of 150 mW/cm2 for a duration of 60 s. In addition, the study assessed the binding affinity of Emodin with GtfB and SlpA proteins using in silico molecular docking. Eventually, the study examined the generation of endogenous reactive oxygen species (ROS) and changes in the gene expression levels of gelE and sprE. RESULTS: The study found that using Emo@PDL-Exo-mediated aPDT resulted in a significant decrease in L. acidophilus and S. mutans by 4.90 ± 0.36 and 5.07 log10 CFU/mL, respectively (P < 0.05). The study found that using Emo@PDL-Exo for aPDT significantly reduced L. acidophilus and S. mutans biofilms by 44.7% and 50.4%, respectively, compared to untreated biofilms in the control group (P < 0.05). Additionally, the metabolic activity of L. acidophilus and S. mutans decreased by 58.3% and 71.2%, respectively (P < 0.05). The molecular docking analysis showed strong binding affinities of Emodin with SlpA and GtfB proteins, with docking scores of -7.4 and -8.2 kcal/mol, respectively. The study also found that the aPDT using Emo@PDL-Exo group resulted in the most significant reduction in gene expression of slpA and gtfB, with a decrease of 4.2- and 5.6-folds, respectively, compared to the control group (P < 0.05), likely due to the increased generation of endogenous ROS. DISCUSSION: The study showed that aPDT using Emo@PDL-Exo can effectively reduce the cell viability, biofilm activity, and metabolic potency of S. mutans and L. acidophilus. aPDT also significantly reduced the expression levels of gtfB and slpA mRNA due to the increased endogenous ROS generation. The findings suggest that Emo@PDL-Exo-mediated aPDT could be a promising antimicrobial approach against cariogenic microorganisms.

Antimicrobial activity of D-amino acid in combination with photo-sonoactivated hypericin nanoparticles against Acinetobacter baumannii.

BACKGROUND: The emergence of multidrug-resistant Acinetobacter baumannii strains is increasing worldwide. To overcome these life-threatening infections, the development of new treatment approaches is critical. For this purpose, this study was conducted to determine the antimicrobial photo-sonodynamic therapy (aPSDT) using hypericin nanoparticles (HypNP) in combination with D-Tryptophan (D-Trp) against A. baumannii. MATERIALS AND METHODS: HypNP was synthesized and characterized, followed by the determination of the fractional inhibitory concentration (FIC) index of HypNP and D-Trp by checkerboard assay. Next, the antimicrobial and anti-biofilm potential of HypNP@D-Trp-mediated aPSDT against A. baumannii was evaluated. Finally, the anti-virulence activity of aPSDT using HypNP@D-Trp was accessed following the characterization of HypNP@D-Trp interaction with AbaI using in silico virtual screening and molecular docking. RESULTS: A synergistic activity in the combination of HypNP and D-Trp against A. baumannii was observed with a FIC index value of 0.5. There was a 5.10 log10 CFU/mL reduction in the cell viability of A. baumannii when the bacterial cells were treated with 1/2 × MIC of HypNP@D-Trp and subsequently exposed to ultrasound waves and blue light (P < 0.05). Moreover, a significant biofilm degradation effect on biofilm-associated cells of A. baumannii was observed after treatment with aPSDT using 2 × MIC of HypNP@D-Trp in comparison with the control groups (P < 0.05). According to the molecular docking analysis of the protein-ligand complex, Hyp with a high affinity for AbaI showed a binding affinity of - 9.41 kcal/mol. Also, the expression level of abaI gene was significantly downregulated by 10.32-fold in A. baumannii treated with aPSDT as comprised with the control group (P < 0.05). CONCLUSIONS: It can be concluded that HypNP@D-Trp-mediated aPSDT can be considered a promising strategy to overcome the infections caused by A. baumannii by reducing the growth of bacterial biofilm and decreasing the expression of abaI as a gene involved in A. baumannii biofilm formation.

Antimicrobial photodynamic therapy: modern technology in the treatment of wound infections in patients with burns.

BACKGROUND: Staphylococcus aureus is one of the most frequently isolated microorganisms from burn wounds. Antimicrobial photodynamic therapy (aPDT) is a new strategy that may improve antimicrobial treatment. METHOD: This study evaluated three meticillin-resistant Staphylococcus aureus (MRSA) and three meticillin-sensitive Staphylococcus aureus (MSSA) clinical isolates, which produced a biofilm with 0.1mg/ml Toluidine Blue O (TBO) (Sigma-Aldrich, Germany) with an energy density of 45J/cm2 and 90J/cm2, for MRSA and MSSA, respectively. The antibiofilm potential of aPDT with TBO was analysed using crystal violet assays and scanning electron microscopy. RESULTS: TBO-aPDT significantly degraded the biofilm formed by MRSA and MSSA clinical isolates (p<0.05). CONCLUSION: Our results indicated that aPDT is an effective approach to combat bacterial biofilms associated with burn wound infection. aPDT could provide a supplemental to the treatment of wound and tissue infection, and patients with burns may benefit from combined treatments.

Theranostic nanoplatforms of emodin-chitosan with blue laser light on enhancing the anti-biofilm activity of photodynamic therapy against Streptococcus mutans biofilms on the enamel surface.

BACKGROUND: Combining photosensitizer and light irradiation, named antimicrobial photodynamic therapy (aPDT) is an adjuvant therapy for eliminating microbial biofilms. This ex vivo study evaluates the effect of anti-biofilm activity of aPDT based on emodin-chitosan nanoparticles (Emo-CS-NPs) plus blue laser light against Streptococcus mutans biofilm on the enamel surface. MATERIALS: After determination of the fractional inhibitory concentration index of Emo and CS by checkerboard array assay, Emo-CS-NPs were synthesized and characterized. Following treatment of pre-formed S. mutans biofilms on the enamel slabs, cellular uptake of Emo-CS-NPs and intracellular reactive oxygen species (ROS) production were determined. The anti-biofilm and anti-metabolic activities of aPDT were investigated. Eventually, lactic acid production capacity, concentrations of S. mutans extracellular DNA (eDNA) levels, and expression of the gene involved in the biofilm formation (gtfB) were evaluated. RESULTS: The maximum uptake of Emo-CS-NPs occurs in an incubation time of 5 min. When irradiated, Emo-CS-NPs were photoactivated, generating ROS, and led to a decrease in the cell viability and metabolic activity of S. mutans significantly (P < 0.05). S. mutans eDNA and lactic acid production outcomes indicated that Emo-CS-NPs-mediated aPDT led to a significant reduction of eDNA levels (48%) and lactic acid production (72.4%) compared to the control group (P < 0.05). In addition, gtfB mRNA expression in S. mutans was downregulated (7.8-fold) after aPDT in comparison with the control group (P < 0.05). CONCLUSIONS: Our data support that, aPDT using Emo-CS-NPs revealed the highest cellular uptake and ROS generation. Emo-CS-NPs based aPDT could inhibit significantly biofilm formation and reduce effectively virulence potency of S. mutans; thus, it could be an adjuvant therapy against dental caries.

Contribution of antimicrobial photo-sonodynamic therapy in wound healing: an in vivo effect of curcumin-nisin-based poly (L-lactic acid) nanoparticle on Acinetobacter baumannii biofilms.

BACKGROUND: The biofilm-forming ability of Acinetobacter baumannii in the burn wound is clinically problematic due to the development of antibiotic-resistant characteristics, leading to new approaches for treatment being needed. In this study, antimicrobial photo-sonodynamic therapy (aPSDT) was used to assess the anti-biofilm efficacy and wound healing activity in mice with established A. baumannii infections. METHODS: Following synthesis and confirmation of Curcumin-Nisin-based poly (L-lactic acid) nanoparticle (CurNisNp), its cytotoxic and release times were evaluated. After determination of the sub-significant reduction (SSR) doses of CurNisNp, irradiation time of light, and ultrasound intensity against A. baumannii, anti-biofilm activity and the intracellular reactive oxygen species (ROS) generation were evaluated. The antibacterial and anti-virulence effects, as well as, histopathological examination of the burn wound sites of treated mice by CurNisNp-mediated aPSDTSSR were assessed and compared with silver sulfadiazine (SSD) as the standard treatment group. RESULTS: The results showed that non-cytotoxic CurNisNp has a homogeneous surface and a sphere-shaped vesicle with continuous release until the 14th day. The dose-dependent reduction in cell viability of A. baumannii was achieved by increasing the concentrations of CurNisNp, irradiation time of light, and ultrasound intensity. There was a time-dependent reduction in biofilm growth, changes in gene expression, and promotion in wound healing by the acceleration of skin re-epithelialization in mice. Not only there was no significant difference between aPSDTSSR and SSD groups in antibacterial and anti-virulence activities, but also wound healing and re-epithelialization occurred more efficiently in aPSDTSSR than in the SSD group. CONCLUSIONS: In conclusion, CurNisNp-mediated aPSDT might be a promising complementary approach to treat burn wound infections.

Antimicrobial and anti-biofilm potencies of dermcidin-derived peptide DCD-1L against Acinetobacter baumannii: an in vivo wound healing model.

BACKGROUND: The global emergence of Acinetobacter baumannii resistance to most conventional antibiotics presents a major therapeutic challenge and necessitates the discovery of new antibacterial agents. The purpose of this study was to investigate in vitro and in vivo anti-biofilm potency of dermcidin-1L (DCD-1L) against extensively drug-resistant (XDR)-, pandrug-resistant (PDR)-, and ATCC19606-A. baumannii. METHODS: After determination of minimum inhibitory concentration (MIC) of DCD-1L, in vitro anti-adhesive and anti-biofilm activities of DCD-1L were evaluated. Cytotoxicity, hemolytic activity, and the effect of DCD-1L treatment on the expression of various biofilm-associated genes were determined. The inhibitory effect of DCD-1L on biofilm formation in the model of catheter-associated infection, as well as, histopathological examination of the burn wound sites of mice treated with DCD-1L were assessed. RESULTS: The bacterial adhesion and biofilm formation in all A. baumannii isolates were inhibited at 2 × , 4 × , and 8 × MIC of DCD-1L, while only 8 × MIC of DCD-1L was able to destroy the pre-formed biofilm in vitro. Also, reduce the expression of genes involved in biofilm formation was observed following DCD-1L treatment. DCD-1L without cytotoxic and hemolytic activities significantly reduced the biofilm formation in the model of catheter-associated infection. In vivo results showed that the count of A. baumannii in infected wounds was significantly decreased and the promotion in wound healing by the acceleration of skin re-epithelialization in mice was observed following treatment with 8 × MIC of DCD-1L. CONCLUSIONS: Results of this study demonstrated that DCD-1L can inhibit bacterial attachment and biofilm formation and prevent the onset of infection. Taking these properties together, DCD-1L appears as a promising candidate for antimicrobial and anti-biofilm drug development.

Quorum quenching of Streptococcus mutans via the nano-quercetin-based antimicrobial photodynamic therapy as a potential target for cariogenic biofilm.

BACKGROUND: Quorum sensing (QS) system can regulate the expression of virulence factors and biofilm formation in Streptococcus mutans. Antimicrobial photodynamic therapy (aPDT) inhibits quorum quenching (QQ), and can be used to prevent microbial biofilm. We thereby aimed to evaluate the anti-biofilm potency and anti-metabolic activity of nano-quercetin (N-QCT)-mediated aPDT against S. mutans. Also, in silico evaluation of the inhibitory effect of N-QCT on the competence-stimulating peptide (CSP) of S. mutans was performed to elucidate the impact of aPDT on various QS-regulated genes. METHODS: Cytotoxicity and intracellular reactive oxygen species (ROS) generation were assessed following synthesis and confirmation of N-QCT. Subsequently, the minimum biofilm inhibitory concentration (MBIC) of N-QCT against S. mutans and anti-biofilm effects of aPDT were assessed using colorimetric assay and plate counting. Molecular modeling and docking analysis were performed to confirm the connection of QCT to CSP. The metabolic activity of S. mutans and the expression level of various genes involved in QS were evaluated by flow cytometry and reverse transcription quantitative real-time PCR, respectively. RESULTS: Successful synthesis of non-toxic N-QCT was confirmed through several characterization tests. The MBIC value of N-QCT against S. mutans was 128 µg/mL. Similar to the crystal violet staining, the results log10 CFU/mL showed a significant degradation of preformed biofilms in the group treated with aPDT compared to the control group (P < 0.05). Following aPDT, metabolic activity of S. mutans also decreased by 85.7% (1/2 × MBIC of N-QCT) and 77.3% (1/4 × MBIC of N-QCT), as compared to the control values (P < 0.05). In silico analysis showed that the QCT molecule was located in the site formed by polypeptide helices of CSP. The relative expression levels of the virulence genes were significantly decreased in the presence of N-QCT-mediated aPDT (P < 0.05). CONCLUSIONS: The combination of N-QCT with blue laser as a QQ-strategy leads to maximum ROS generation, disrupts the microbial biofilm of S. mutans, reduces metabolic activity, and downregulates the expression of genes involved in the QS pathway by targeting genes of the QS signaling system of S. mutans.

Anti-leishmanial effects of resveratrol and resveratrol nanoemulsion on Leishmania major.

BACKGROUND: Leishmaniasis is a vector-borne disease that is endemic in the tropical and sub-tropical areas of the world. Low efficacy and high cytotoxicity of the current treatment regimens for leishmaniasis is one of the most important health problems. In this experimental study, anti-leishmanial effects of different concentrations of resveratrol and resveratrol nano-emulsion (RNE) were assessed. METHODS: RNE was prepared using the probe ultra-sonication method. The cytotoxicity was evaluated using the MTT technique on the L929 cell line. The anti-leishmanial activities on promastigotes of leishmania were assessed using vital staining and infected BALB/c mice were used to assess the in vivo anti-leishmanial effects. RESULTS: In vitro and in vivo assays revealed that all concentrations of resveratrol and RNE had valuable inhibitory effects against Leishmania major in comparison to the control group (P < 0.05). The half maximal inhibitory concentration (IC50) values were calculated as 16.23 and 35.71 µg/mL for resveratrol and RNE, respectively. Resveratrol and RNE showed no cytotoxicity against the L929 cell line. CONCLUSIONS: According to the potent in vitro and in vivo anti-leishmanial activity of RNE at low concentration against L. major, we suggest that it could be a promising anti-leishmanial therapeutic against L. major in the future.

Molecular Modeling and Simulation Analysis of Antimicrobial Photodynamic Therapy Potential for Control of COVID-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can enter the host cells by binding the viral surface spike glycoprotein (SG) to angiotensin-converting enzyme 2. Since antiviral photodynamic therapy (aPDT) has been described as a new method for inhibiting viral infections, it is important to evaluate whether it can be used as a photoactivated disinfectant to control COVID-19. In this in silico study, SARS-CoV-2-SG was selected as a novel target for curcumin as a photosensitizer during aPDT to exploit its physicochemical properties, molecular modeling, hierarchical nature of protein structure, and functional analysis using several bioinformatics tools and biological databases. The results of a detailed computational investigation revealed that SARS-CoV-2-SG is most similar to 6VXX_A, with 100% query cover and identity. The predicted structure of SARS-CoV-2-SG displayed that it is a protein with a positive charge and random coil dominates other secondary structures located outside the viral cell. The protein-protein interaction network showed that SARS-CoV-2-SG interacted with ten potential interacting partners. In addition, primary screening of binding modes through molecular docking showed that curcumin desires to bind and interact with residues of SARS-CoV-2-SG as the main site to enhance the yield of aPDT. Overall, the computer simulation reveals that SARS-CoV-2-SG can be a suitable target site for interaction with curcumin during aPDT.

Photoactivation of Curcumin Doped Poly-Lactic-Co-Glycolic Acid Nanoparticles in Rat Model with Fixed Orthodontic Appliances.

This study aimed to evaluate the antimicrobial effect of curcumin doped poly-lactic-co-glycolic acid nanoparticles (Cur-PLGA-Nps)-mediated antimicrobial photodynamic therapy (aPDT), as well as the probiotics on S. mutans in rats with fixed orthodontic appliances. Orthodontic appliances were ligated to the right maxillary of the rats. After the oral colonization of S. mutans, the rats were then treated in four groups including Cur-PLGA-Nps, light-emitting diode, Cur-PLGA-Nps-mediated aPDT, and probiotic (Lactobacillus acidophilus). After that, the S. mutans counts and the gtfB gene expression of S. mutans were determined on days 4, 7, 15, and 30. Probiotic and Cur-PLGA-Nps-mediated aPDT groups significantly reduced the count of S. mutans in a time-dependent manner (P < 0.05). So, probiotics and Cur-PLGA-Nps-mediated aPDT were able to reduce S. mutans more than other groups on the 30th day. Also, there was no considerable difference between Cur-PLGA-Nps-mediated aPDT and probiotic groups in bacterial growth inhibition. The expression level of gtfB gene was significantly downregulated at all-time intervals after exposure to Cur-PLGA-Nps-mediated aPDT compared with untreated bacteria (P < 0.05). According to the results, simultaneous use of Cur-PLGA-Nps-mediated aPDT and probiotic therapeutic approaches can be suggested to increase effectiveness.

Antimicrobial effects and mechanical properties of poly(methyl methacrylate) as an orthodontic acrylic resin containing Curcumin-Nisin-poly(L-lactic acid) nanoparticle: an in vitro study.

BACKGROUND: The porous surface of acrylic orthodontic removable appliances creates a niche for microbial plaque accumulation, and changes the oral flora by raising cariogenic bacteria including Streptococcus mutans. In this study, we evaluated the mechanical properties and antimicrobial activities of incorporating different concentrations of Curcumin-Nisin-poly(L-lactic acid) nanoparticle (CurNisNps) into orthodontic acrylic resin against Streptococcus mutans and Candida albicans. METHODS: Following synthesis and characterization of CurNisNps, acrylic resin specimens with different concentrations of CurNisNps (0, 1, 2, 5, and 10% w/w) were fabricated. Flexural strength values, antimicrobial effects, anti-biofilm potential, and anti-metabolic activity against S. mutans and C. albicans were assessed at different time intervals. Also, the expression of the virulence-factor-related genes of S. mutans and C. albicans was assessed by quantitative real-time polymerase chain reaction following treatment with CurNisNps. RESULTS: Acrylic resin containing 10% CurNisNps (30.76 ± 3.91 MPa) showed flexural failure in comparison with acrylic resin specimens without CurNisNps (50.67 ± 1.82 MPa) as the control group (P < 0.05). There was no significant decrease in the flexural strength values in samples containing 1, 2, and 5% of CurNisNps in comparison to the control group (P > 0.05). Acrylic resin with 5% CurNisNps showed the highest concentration of CurNisNps and clinically accepted flexural strength value (14.89 ± 3.26 MPa, P < 0.05) simultaneously. In the disc agar diffusion assay, 5% CurNisNps showed a high level of inhibitory activity for the test microorganisms. The reduction of growth inhibition zones of the different concentrations of CurNisNps against test microorganisms was positively associated with the time, in such a way that it was reduced significantly after 60 days. The anti-biofilm and anti-metabolic activities of acrylic resin specimens containing a 5% concentration of CurNisNps against S. mutans and C. albicans could significantly decrease the expression levels of gtfB (6.8-fold) and HWP (3.4-fold) in S. mutans and C. albicans, respectively. CONCLUSIONS: Our data support that 5% (w/w) of CurNisNps can serve as an excellent orthodontic acrylic resin additive against S. mutans and C. albicans biofilm without adverse effects on its mechanical property.

Enhanced reduction of polymicrobial biofilms on the orthodontic brackets and enamel surface remineralization using zeolite-zinc oxide nanoparticles-based antimicrobial photodynamic therapy.

The aim of this study was to evaluate the anti-biofilm and anti-metabolic activities of zeolite-zinc oxide nanoparticles (Zeo/ZnONPs)-based antimicrobial photodynamic therapy (aPDT) against pre-formed polymicrobial biofilms on the orthodontic brackets, as well as, assess the remineralization efficacy on polymicrobial biofilms induced enamel lesions. Following synthesis and characterization of Zeo/ZnONPs, cell cytotoxicity, hemolytic effect, and intracellular reactive oxygen species (ROS) production were determined. The anti-biofilm and anti-metabolic activities of aPDT using different concentrations of Zeo/ZnONPs were investigated. Microhardness tester and DIAGNOdent Pen were used to evaluate the changes of remineralization degree on the treated enamel slabs duration 1 and 3 months. No significant cytotoxicity and erythrocyte hemolysis were observed in treated cells with Zeo/ZnONPs. When irradiated, suggesting that the Zeo/ZnONPs were photoactivated, generating ROS and leading to reduce dose-dependently the cell viability and metabolic activity of polymicrobial biofilms. Also, the enamel surface microhardness value of exposed enamel showed a steady increase with the concentration of Zeo/ZnONPs. No statistically significant differences were shown between aPDT and sodium fluoride varnish as the control group. Overall, Zeo/ZnONPs-based aPDT with the greatest remineralization efficacy of enamel surface can be used as an anti-biofilm therapeutic method, which is involved with their potent ability to produce ROS.

Photobiomodulation and Antiviral Photodynamic Therapy in COVID-19 Management.

Coronavirus disease 2019 (COVID-19) has shocked the world by its spread and contagiousness. There is no approved vaccine and no proven treatment for this infection. Some potential treatments that have already been associated with antiviral and anti-inflammatory effects are under investigation. Photobiomodulation therapy (PBMT) is a photon-based therapy that uses light to mediate a variety of metabolic, analgesic, anti-inflammatory, and immunomodulatory effects. Antiviral photodynamic therapy (aPDT) is a branch of photodynamic therapy based on the reaction between a photosensitizing agent and a light source in the presence of oxygen, which can produce oxidative and free radical agents to damage the viral structures such as proteins and nucleic acids. This chapter aims to discuss the potential therapeutic benefit of PBMT and aPDT in the context of the novel coronavirus. Studies indicate that PBMT and aPDT could be useful in many viral and bacterial pulmonary complications like influenza, SARS-CoV, and MERS, but we found no direct study on SARS-CoV-2. With a combination of PBMT and aPDT, we may be able to combat COVID-19 with minimal interference with pharmaceutical agents. It might improve the efficacy of PBMT and aPDT by using monoclonal antibodies and preparing new photosensitizers at the nanoscale that target the lung tissue specifically. More animal and human studies would need to take place to reach an effective protocol. This chapter would encourage other scientists to work on this new platform.

Evaluation of the Antimicrobial Effect of Mineral Trioxide Aggregate Mixed with Fluorohydroxyapatite against E. faecalis In Vitro.

Enterococcus faecalis is the dominant microorganism in chronic apical periodontitis. It is more resistant to local antiseptic agents than other endodontic microorganisms. Currently, mineral trioxide aggregate (MTA) is considered as an ideal material in many endodontic procedures. Some studies have shown that MTA has good antibacterial activity against E. faecalis. However, some studies have investigated the effect of incorporating some materials into MTA on its antibacterial activity against E. faecalis. No study has evaluated the effect of incorporating fluorohydroxyapatite nanoparticles (nano-FHA) on the antimicrobial activity of MTA. Therefore, the present study evaluated the antimicrobial effect of MTA mixed with nano-FHA on E. faecalis in vitro. The study was carried out on 18 samples in three groups: pure MTA, MTA mixed with 10 wt% of nano-FHA, and MTA mixed with 15 wt% of nan-FHA. The effect of nano-FHA on the antibacterial activity of MTA on E. faecalis was evaluated by evaluating the growth inhibition zone around each sample. The antimicrobial effect of samples on inhibiting E. faecalis biofilm formation and inhibiting microbial growth of E. faecalis in the planktonic phase was evaluated by disk agar diffusion (DAD), biofilm inhibition assay (BIA), and direct contact assay (DCA) tests, respectively. All the above tests were analyzed after 24 and 72 hours. Factorial designs were used for statistical analyses. Tukey tests were used for two-by-two comparisons. All the statistical analyses were carried out with SPSS 26. DAD results showed no formation of the growth inhibition zone in all the samples after 24 and 72 hours. The microbial colony counts in the BIA and DCA tests in the groups modified with FHA nanoparticles were significantly lower than the pure MTA group (P < 0.05). The microbial colony counts increased in all the groups over time (P < 0.05). Incorporating nano-FHA into MTA improved the antimicrobial activity of MTA against E. faecalis compared to pure MTA. The highest antimicrobial activity was achieved after incorporating 15 wt% of nano-FHA into MTA at the 72-hour interval.

Therapeutic applications of nucleic acid aptamers in microbial infections.

Today, the treatment of bacterial infections is a major challenge, due to growing rate of multidrug-resistant bacteria, complication of treatment and increased healthcare costs. Moreover, new treatments for bacterial infections are limited. Oligonucleotide aptamers are single stranded DNAs or RNAs with target-selective high-affinity feature, which considered as nucleic acid-based affinity ligands, replacing monoclonal antibodies. The aptamer-based systems have been found to be talented tools in the treatment of microbial infections, regarding their promising anti-biofilm and antimicrobial activities; they can reduce or inhibit the effects of bacterial toxins, and inhibit pathogen invasion to immune cell, as well as they can be used in drug delivery systems. The focus of this review is on the therapeutic applications of aptamers in infections. In this regard, an introduction of infections and related challenges were presented, first. Then, aptamer definition and selection, with a brief history of aptamers development against various pathogens and toxins were reviewed. Diverse strategies of aptamer application in drug delivery, as well as, the effect of aptamers on the immune system, as the main natural agents of human defense against pathogens, were also discussed. Finally, the future trends in clinical applications of this technology were discussed.

Effect of Addition of Nano-TiO2, Nano-SiO2, and a Combination of Both, on Antimicrobial Activity of an Orthodontic Composite.

AIM: This study aimed to assess the effect of addition of nano-titanium oxide (nano-TiO2), nano-silicon dioxide (nano-SiO2), and a combination of both, on antimicrobial activity of an orthodontic composite. MATERIALS AND METHODS: Molds measuring 0.64 × 0.5 mm were used for the fabrication of composite disks. For this purpose, 0.5% and 1% nano-TiO2, nano-SiO2, and a combination of both (0.5% nano-TiO2 and 0.5% nano-SiO2), were mixed with Transbond XT composite (3M Unitek). A total of 180 composite disks were fabricated for eluted component, disk agar diffusion (DAD), and biofilm inhibition tests. The colony counts of Streptococcus mutans (S. mutans), Streptococcus sanguinis (S. sanguinis), and Lactobacillus acidophilus (L. acidophilus) and the diameters of growth inhibition zones were measured at 3, 15, and 30 days after exposure to the materials. Data were analyzed using one-way ANOVA and a post hoc test. RESULTS: None of the nano-TiO2 and nano-SiO2 concentrations had any significant effect on the growth inhibition zone. All tested concentrations of nano-TiO2 and nano-SiO2 decreased the colony count of all bacteria. The composite sample containing both nano-TiO2 and nano-SiO2 had the greatest efficacy for reduction of S. mutans and S. sanguinis colony counts at all three time points. Also, 1% nano-TiO2 and 1% nano-SiO2 had similar effects on L. acidophilus in eluted component test. CONCLUSION: Addition of TiO2 and SiO2 nanoparticles conferred antimicrobial property to the tested orthodontic composite. CLINICAL SIGNIFICANCE: Using orthodontic composite containing nonoparticles with antibacterial activity may prevent dental caries.

Biofilm formation and antibiotic resistance in meticillin-resistant and meticillin-sensitive Staphylococcus aureus isolated from burns.

OBJECTIVE: To investigate the relationship between biofilm formation and antibiotic resistance patterns in meticillin-resistant and meticillin-sensitive Staphylococcus aureus, isolated from burns. METHODS: In a cross-sectional study, pus/wound swab samples were obtained from burns. Presence of Staphylococcus aureus was confirmed, and biofilm formation-related icaABCDR and eta, etb genes were detected by polymerase chain reaction. Biofilm formation assay was assessed using the microtiter plate method. Antibiotic resistance was performed using the disk diffusion method and minimum inhibitory concentration. RESULTS: A total of 95 patients with burns were recruited. Of the 95 wounds swabbed, Staphylococcus aureus was identified in 50 (62.5%), and 47 (94%) isolates capable of producing biofilm. Biofilm production levels were classed as 'strong' (n=29; 58%), 'moderate' (n=11; 22%), 'weak' (n=7; 14%) and 'non-biofilm forming' (n=3; 6%). There was an almost even split between isolates identified as meticillin-resistant Staphylococcus aureus (MRSA), n=24 (48%), and meticillin-sensitive Staphylococcus aureus (MSSA) n=26 (52%). The prevalence of the icaA, icaB, icaC, icaD and icaR genes among the studied isolates were 96%, 80%, 80%, 96% and 84%, respectively. The prevalence of eta and etb genes in isolates were 84% and 92%, respectively. CONCLUSION: Biofilm producing isolates of Staphylococcus aureus showed greater multidrug resistance than non-biofilm producers. In our study, a high rate of biofilm formation and antimicrobial drug resistance was seen. Our results highlight the alarming levels of antimicrobial resistance among MRSA strains and important data about the prevalence of eta and etb genes in Staphylococcus aureus strains isolated from burn patients in this study.

Effect of 5 Popular Disinfection Methods on Microflora of Laboratory: Customized Implant Abutments.

PURPOSE: To compare the efficacy of 5 different decontamination methods of titanium abutments and to assess their possible effects on surface roughness of titanium. MATERIALS AND METHODS: Micrococcus luteus, Acinetobacter baumannii, Enterococcus faecalis, and Candida albicans were cocultured to form a multispecies biofilm on 18 titanium discs. In another group, Bacillus subtilis, a spore-forming species, was cultivated on another set of 18 titanium discs. Each group was further divided into 5 test groups: high-pressure steam cleaning (4 Mpa, 5 seconds), NaOCl (1% active chlorine, 5 minutes), H2O2 (3%, 5 minutes), GaAlAs laser (810 nm, CW, 1 W, 400-µm fiber, 1-mm distance, 1 minute), Er:YAG laser (2940 nm, pulse mode, 100 mJ, 10 Hz, 230-µm noncontact handpiece, 4-mm distance, 50/50% air/water, 1 minute), and a control group of no treatment. After each decontamination procedure, the remaining microbial load was reported as colony-forming unit/disc. To evaluate the effect of each treatment on titanium discs, surface roughness parameters including Sa, Sq, Ssk, Sku, Sal, and Sdr were measured at 6 points of each disc using an atomic force microscope. RESULTS: Complete disinfection was achieved using high-pressure steam, NaOCl, H2O2, and Er:YAG laser. GaAlAs laser was able to reduce microbial count over 90%. Sa and Sq parameters were only increased significantly in diluted NaOCl group in comparison with control group, whereas Sdr was increased significantly in both absolute and diluted NaOCl groups. CONCLUSION: All the methods could decontaminate machined titanium surfaces, although complete microbial elimination was not achieved by diode laser. None of the treatments altered surface roughness significantly, except for sodium hypochlorite (NaOCl).

Association of virulence gene expression with colistin-resistance in Acinetobacter baumannii: analysis of genotype, antimicrobial susceptibility, and biofilm formation.

BACKGROUND: Acinetobacter baumannii causes difficult-to-treat nosocomial infections, which often lead to morbidity due to the development of antimicrobial drug resistance and expression of virulence genes. Data regarding the association of resistance to colistin, a last treatment option, and the virulence gene expression of A. baumannii is scarce. METHODS: We evaluated the MLVA genotype, antimicrobial resistance, and biofilm formation of 100 A. baumannii isolates from burn patients, and further compared the in vitro and in vivo expression of four virulence genes among five colistin-resistant A. baumannii (Cst-R-AB) isolates. Five Cst-R-AB isolates were tested; one from the present study, and four isolated previously. RESULTS: Our results showed that reduced expression of recA, along with increased in vivo expression of lpsB, dnaK, and blsA; are associated with colistin resistance among Cst-R-AB isolates. Differences in virulence gene expressions among Cst-R-AB isolates, may in part explain common discrepant in vitro vs. in vivo susceptibility data during treatment of infections caused by Cst-R-AB. CONCLUSIONS: Our findings highlight the intricate relationship between colistin-resistance and virulence among A. baumannii isolates, and underscore the importance of examining the interactions between virulence and antimicrobial resistance toward efforts to control the spread of multidrug-resistant A. baumannii (MDR-AB) isolates, and also to reduce disease severity in burn patients with MDR-AB infection.