Synthesis and Characterization of Paclitaxel-Loaded Silver Nanoparticles: Evaluation of Cytotoxic Effects and Antimicrobial Activity.

Carrier system therapies based on combining cancer drugs with nanoparticles have been reported to control tumor growth and significantly reduce the side effects of cancer drugs. We thought that paclitaxel-loaded silver nanoparticles (AgNPs-PTX) were the right carrier to target cancer cells. We also carried out antimicrobial activity experiments as systems formed with nanoparticles have been shown to have antimicrobial activity. In our study, we used easy-to-synthesize and low-cost silver nanoparticles (AgNPs) with biocatalytic and photocatalytic advantages as drug carriers. We investigated the antiproliferative activities of silver nanoparticles synthesized by adding paclitaxel on MCF-7 (breast adenocarcinoma cell line), A549 (lung carcinoma cell line), C6 (brain glioma cell line) cells, and healthy WI-38 (fibroblast normal cell line) cell lines and their antimicrobial activities on 10 different microorganisms. The synthesized AgNPs and AgNPs-PTX were characterized by dynamic light scattering (DLS), scanning transmission electron microscopy, UV-visible spectroscopy, Fourier transform infrared spectroscopy, and X-ray spectroscopy. The nanoparticles were spherical in shape, with AgNPs ranging in size from 2.32 to 5.6 nm and AgNPs-PTXs from 24.36 to 58.77 nm. AgNPs demonstrated well stability of -47.3 mV, and AgNPs-PTX showed good stability of -25.4 mV. The antiproliferative effects of the synthesized nanoparticles were determined by XTT (tetrazolium dye; 2,3-bis-(2-methoxy-4-nitro-5-sulfenyl)-(2H)-tetrazolium-5-carboxanilide), and the proapoptotic effects were determined by annexin V/propidium iodide (PI) staining. The effect of AgNPs-PTX was more effective, and anticancer activity was higher than PTX in all cell lines. When selectivity indices were calculated, AgNPs-PTX was more selective in the A549 cell line (SI value 6.53 µg/mL). AgNPs-PTX was determined to increase apoptosis cells by inducing DNA fragmentation. To determine the antimicrobial activity, the MIC (minimum inhibitory concentration) test was performed using 8 different bacteria and 2 different fungi. Seven of the 10 microorganisms tested exhibited high antimicrobial activity according to the MIC ≤100 µg/mL standard, reaching MIC values below 100 µg/mL and 100 µg/mL for both AgNPs and AgNPs-PTX compared to reference sources. Compared to standard antibiotics, AgNPs-PTX was highly effective against 4 microorganisms.

Research into biocompatibility and cytotoxicity of daptomycin, gentamicin, vancomycin and teicoplanin antibiotics at common doses added to bone cement.

OBJECTIVES: This study aims to investigate the biocompatibility and cytotoxicity of daptomycin, gentamicin, vancomycin and teicoplanin at commonly-used dose intervals added to polymethylmethacrylate (PMMA) in vitro. MATERIALS AND METHODS: This prospective study was conducted between February 2016 and June 2016. Antibiotics were added to PMMA at doses frequently used in clinical practice. The antibiotic doses added were teicoplanin (2 g, 3 g, 4 g), gentamicin (0.5 g, 0.75 g, 1 g), daptomycin (0.5 g.) and vancomycin (2 g, 3 g, 4 g). Standard cement balls (10 mm) were created. Activated L929 mouse fibroblast cell culture was used for incubation. Agar diffusion, Cell Proliferation Kit II (XTT) test and electron microscope investigations were performed to examine biocompatibility and cytotoxicity. RESULTS: In the cytotoxicity test, teicoplanin at 4 g and daptomycin at 0.5 g doses were observed to cause reductions in viability percentages. The same doses caused 20% and 20-40% cell lysis indices during the agar diffusion test. On electron microscope images, cytotoxic effects in fibroblast cells and involvement with the surface of cement balls were observed. CONCLUSION: Gentamicin, vancomycin and teicoplanin were observed to be non-toxic and biocompatible at commonly-used dose intervals. Teicoplanin at 4 g and daptomycin at 0.5 g doses were identified to be cytotoxic and not biocompatible. When selecting antibiotics to be added to bone cement, care should be taken that the antibiotic is non-toxic and biocompatible.

Amoebicidal activity of benzothiazole on Acanthamoeba castellanii cysts and trophozoites and its cytotoxic potentials.

Acanthamoeba spp. are free-living amoebae isolated from many ecological areas such as swimming pools, dams and lakes, and soil. Granulomatous amebic encephalitis and amebic keratitis, caused by Acanthamoeba spp., usually occurs in chronically ill, debilitated individuals, in immunosuppressed patients and treatment is quite difficult. This study aimed to determine the effect of benzothiazole on trophozoite and cyst forms of Acanthamoeba castellanii (A.castellanii). Axenic cultures of A. castellanii trophozoites and cysts were prepared to test the amoebicidal activity of benzothiazole. The concentrations of benzothiazole in 24-well plates were prepared as 0.08%, 0.04%, 0.02%, 0.01%, 0.005%, and A. castellanii cysts and trophozoites were added to these cultures. Parasites were counted at 0, ½, 1, 3, 6, 12, 24 and 48 h in the cell counter after staining with trypan blue. Cytotoxicity of benzothiazole on the WI-38 human fibroblast cell line was also tested. Between 0.08% and 0.01% concentrations of benzothiazole showed a strong amoebicidal activity at 24 and 48 h. A significant decrease in 0.005% concentration in the number of live trophozoites and cysts was detected between 6 and 48 h. As a result of the cytotoxicity studies, benzothiazole did not show any cytotoxic effect on the WI-38 human fibroblast cell line even at 1% concentration. Benzothiazole could be concluded as a new therapeutic agent against Acanthamoeba. On the other hand, in vivo studies are needed to confirm the efficacy of the biological effect.

Apical extrusion of intracanal biofilm using ProTaper Gold, WaveOne Gold,Twisted File Adaptive, OneShape New Generation and K3XF.

OBJECTIVE: To evaluate the bacterial extrusion during instrumentation with different nickel titanium (NiTi) engine-driven instruments. METHODS: Ninety extracted single-canal human mandibular incisor teeth were inoculated with Enterococcus faecalis to obtain biofilm formation and were randomly divided to 6 groups (n=15). One group served as the control and was not instrumented; the other groups were prepared with ProTaper Gold (PTG; Dentsply Maillefer, Ballaigues, Switzerland), WaveOne Gold (WOG; Dentsply Maillefer), Twisted File Adaptive (TFA; SybronEndo, Orange, CA, USA), One Shape New Generation (OSNG; MicroMega, Besancon, France), and K3XF (SybronEndo) instruments. Bacteria extruded beyond the apical foramen were quantified in colony-forming units per milliliter. The number of colony-forming units in the remaining biofilm was determined for each sample. Data were analyzed using the one-way analysis of variance (ANOVA) and Tukey post-hoc tests. RESULTS: All NiTi instruments resulted in different quantities of bacterial extrusion. The TFA group caused most bacterial extrusion (P<0.05). The PTG and WOG groups caused less bacterial extrusion than the OSNG and K3XF groups (P<0.05), but there was no statistically significant difference between the PTG and WOG groups (P>0.05). CONCLUSION: PTG and WOG are preferable system in terms of successful endodontic treatments. The amount of bacterial extrusion is associated with the metallurgy and design of the instrument used.

Antibacterial Efficacy of Super-Oxidized Water on Enterococcus faecalis Biofilms in Root Canal.

BACKGROUND: The success of endodontic treatment depends on a few crucial factors. One of these factors is the complete chemomechanic preparation of root canal against various bacteria. In particular, the effect of resistant bacteria may cause intense pain with flare-up and formation of periapical lesions. Therefore, the strong effect of irrigants plays an important role in terms of the complete elimination of these bacteria to achieve long-term successful treatment. OBJECTIVES: The aim of this study was to investigate the antibacterial effects of super-oxidized water (SPO) in root canals infected with Enterococcus faecalis biofilms. METHODS: One hundred twenty single-root, premolar teeth were selected. Initially, the teeth were prepared and then disinfected. E. faecalis were inoculated and kept at 37°C for 24 hours in the root canals. The re-inoculation procedure was repeated on the first, fourth, seventh, and tenth days. The infected root canals were divided into one negative (saline) and one positive (sodium hypochlorite) control group and four experimental groups (super-oxidized water: 1, 2, 3, or 5 minutes) (n = 20). Paper points were placed in the root canals to control and evaluate the biofilm formation. Biofilms were counted on blood agar plates, and data was evaluated and statistically analyzed using one-way ANOVA and Tukey's test. RESULTS: Although sodium hypochlorite (NaOCl) showed no statistically significant difference when compared with three and five minutes of SPO irrigation (P > 0.05), NaOCl showed statistically significant differences among all other groups (P < 0.05). CONCLUSIONS: Super-oxidized water indicated a remarkable and similar bactericidal effect to that of traditional NaOCl against E. faecalis biofilms. In terms of successful endodontic treatment approaches, super-oxidized water may be used as an effective irrigation solution in clinics.

Bactericidal effects of various irrigation solutions against staphylococcus aureus in human root canal.

PURPOSE: The aim of the present study was to evaluate and to compare the antibacterial effects of various irrigation solutions against Staphylococcus aureus (S. aureus) in human root canals. MATERIALS AND METHODS: 120 single-root mandibular premolar teeth were selected. The teeth were prepared and sterilized. S. aureus was incubated in the root canals and kept at 37°C for 24h. The infected root canals were divided into one positive (saline) and one negative (sodium hypochlorite) control, and four experimental groups [Ethylene-diaminetetra-aceticacid, Chlorhexidine Gluconate, Super-oxidized water(SPO), Aqueous ozone] (n=20). Flow rate of irrigation was applied with 5 mL/min flow rate for 3 min to ensure standardization among all study groups. Following the irrigation, paper points were placed in the root canals and then transferred in sterile eppendorf. Remaining bacteria were counted on blood agar plates and the data were analyzed using one-way ANOVA and Tukey's test. RESULTS: Although there were statistically significant differences among SPO and other experimental groups (p<0.05), there was no statistically significant difference between SPO and NaOCl (p>0.05). CONCLUSION: Super-oxidized water may be recommended as an alternative irrigation solution instead of NaOCl against S. aureus in root canals.

Antibacterial Efficacy of Aqueous Ozone in Root Canals Infected by Enterococcus faecalis.

BACKGROUND: In endodontics, the elimination of resistant bacteria such as Enterococcus faecalis plays an important role for treatment success in root canals. Therefore, new alternative irrigants (instead of sodium hypochlorite) have been researched to achieve ideal endodontic treatment. OBJECTIVES: The aim of the present study was to evaluate and to compare the antibacterial effect of aqueous ozone with different concentrations and techniques of application (manual and ultrasonic) against E. faecalis in human root canals. PATIENTS AND METHODS: Eighty single-root mandibular premolar teeth were selected, prepared and sterilized. E. faecalis was incubated in the root canals and kept at 37°C for 24 h. The teeth were divided into four main groups each has 20 members: NaOCl (positive control) group; 8 ppm aqueous ozone group; 12 ppm aqueous ozone group; and 16 ppm aqueous ozone group. While half of the specimens were disinfected with aqueous ozone by manual technique, the other half was disinfected with the aqueous ozone by ultrasonic technique. Conventional irrigation technique was simultaneously applied with ultrasonic vibration that was produced by VDW.ULTRA device. The disinfection procedures were performed for 180 s to ensure standardization of all the working groups. Paper points (placed in the root canals before and after the disinfection procedures) were transferred to Eppendorf tubes containing 0.5 mL of brain heart infusion broth. Then, 50 µL of the suspension was inoculated onto broth agar media. Microbial colonies were counted, and the data were evaluated statistically using 2-way analysis of variance (ANOVA) and Tukey tests. RESULTS: Although the antibacterial effect of 16 ppm aqueous ozone using a manual technique had an insufficient effect, its ultrasonic application technique resulted in complete disinfection in the root canals. CONCLUSIONS: The bactericidal activity of high concentration of aqueous ozone combined with ultrasonic application technique showed efficacy similar to that of 5.25% NaOCl in root canals.

Antibacterial effects of two different types of laser and aqueous ozone against Enterococcus faecalis in root canals.

OBJECTIVE: The purpose of this study was to investigate the antibacterial effects of two different types of laser and aqueous ozone in human root canals infected by Enterococcus faecalis. BACKGROUND DATA: Many techniques have been developed to find an alternative to sodium hypochlorite as a disinfection agent for infected root canals. However, no study has evaluated the exactly the same antimicrobial agent with sodium hypochlorite (NaOCl). METHODS: Eighty mandibular premolar teeth with single roots and canals were selected. Following root canal preparation and irrigation, sterilization was performed in an autoclave. E. faecalis was incubated in the root canals and kept at 37°C for 24 h. The teeth contaminated with E. faecalis were divided into one negative control group (NaOCl) and three experimental groups; (Er:YAG laser, KTP laser, and aqueous ozone groups)(n=20). A disinfection procedure was performed for 3 min in order to standardize all groups. After this procedure, the microbial colonies were counted. RESULTS: The results indicated that whereas the NaOCl group exhibited the highest antibacterial effect among all groups, the aqueous ozone showed the highest antibacterial effect among the experimental groups. Whereas a statistically significant difference was noted between the aqueous ozone and laser groups (p<0.05), the difference between the Er:YAG and KTP lasers was not statistically significant (p>0.05). CONCLUSIONS: The results showed that when aqueous ozone was applied with the aim of disinfecting the root canals, it exhibited a higher antibacterial effect than the KTP and Er:YAG lasers. However, the antibacterial effect of the aqueous ozone was insufficient when compared with NaOCl.

The effect of low dose teicoplanin-loaded acrylic bone cement on biocompatibility of bone cement.

Antibiotic-loaded acrylic bone cement (polymethylmethacrylate, PMMA) is used to prevent or treat infection in total joint replacement surgery. The purpose of this study was to investigate biocompatibility and cytotoxicity of the teicoplanin-loaded acrylic bone cement. Cytotoxicity examination of acrylic bone cement balls and 400 mg teicoplanin added acrylic bone cement balls conducted by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. SEM (Scanning electron microscopy) was used to observe adhesion and spreading of cells on surface of the balls. Cytotoxicity examination conducted by MTT assay on acrylic bone cement balls and teicoplanin-added acrylic bone cement balls revealed no cytotoxicity. SEM analysis put forward that cells started to proliferate and adhere on surface of the samples in both groups as a result of 48-hour incubation and that the cell proliferation over acrylic bone cement and teicoplanin-added acrylic bone cement was similar. As a consequence, there was no cytotoxicity in acrylic bone cement and teicoplanin-added acrylic bone cement groups according to results of MTT assay. On the other hand, results of SEM showed that biocompatibility of both groups was similar. In conclusion, teicoplanin-loaded bone cement did not change biocompatibility of bone cement in studied dose.