Purpose: The purpose of this study was to compare the phototoxic effects of blue light exposure on periodontal pathogens in both planktonic and biofilm cultures. Methods: Strains of Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, and Porphyromonas gingivalis, in planktonic or biofilm states, were exposed to visible light at wavelengths of 400.520 nm. A quartz-tungsten-halogen lamp at a power density of $500mW/cm^2$ was used for the light source. Each sample was exposed to 15, 30, 60, 90, or 120 seconds of each bacterial strain in the planktonic or biofilm state. Confocal scanning laser microscopy (CSLM) was used to observe the distribution of live/dead bacterial cells in biofilms. After light exposure, the bacterial killing rates were calculated from colony forming unit (CFU) counts. Results: CLSM images that were obtained from biofilms showed a mixture of dead and live bacterial cells extending to a depth of $30-45{\mu}m$. Obvious differences in the live-to-dead bacterial cell ratio were found in P. gingivalis biofilm according to light exposure time. In the planktonic state, almost all bacteria were killed with 60 seconds of light exposure to F. nucleatum (99.1%) and with 15 seconds to P. gingivalis (100%). In the biofilm state, however, only the CFU of P. gingivalis demonstrated a decreasing tendency with increasing light exposure time, and there was a lower efficacy of phototoxicity to P. gingivalis as biofilm than in the planktonic state. Conclusions: Blue light exposure using a dental halogen curing unit is effective in reducing periodontal pathogens in the planktonic state. It is recommended that an adjunctive exogenous photosensitizer be used and that pathogens be exposed to visible light for clinical antimicrobial periodontal therapy.
In this study we determined fertilization processes and developmental ability of porcine oocytes following injection of round spermatid in the presence of artificial activation. Electrical stimulation at 3 h before spermatid injection significantly increased the incidence of normal fertilization as compared to those following injection without stimulation or with stimulation immediately after injection. The incidences of two pronuclear formation and apposition were not different in oocytes between following intracytoplasmic spermatid and spermatid nucleus injection. Indirect immunocytochemistry and laser scanning confocal microscopy study revealed that micro tubules were organized from the oocyte cortex following round spermatid injection, and this seemed to move both male and female pronuclei into the oocyte center. Paternal mitochondria which are introduced with spermatid have been observed up to 4-cell. Our study indicated that either round spermatid or it's nucleus can be used to produce viable bovine embryos by injection into unfertilized oocytes.
Kim, Hyungjun;Park, Soyoung;Jeong, Taesung;Kim, Shin
Journal of the korean academy of Pediatric Dentistry
/
v.46
no.4
/
pp.382-391
/
2019
This study was aimed to assess the new trial for minimal cavity preparation in composite restoration combined with resin infiltration, focusing at application sequence. 32 human primary molars with early carious lesions around small cavity were selected and randomly divided into two groups, according to the sequence of cavity preparation (P), composite filling (F) and resin infiltration (I) as IPF and PFI group. Each group was assessed about amount of tooth reduction, features of resin infiltration, and marginal leakage around restoration. Amount of tooth reduction evaluated using micro-CT was decreased compared with the original lesion size in both groups. Features of resin infiltration were verified under confocal laser scanning microscopy. In both groups, infiltrant resin was found on all around the composite and maintained in spite of extent of decalcification even after artificial caries induction. Marginal micro leakage assessed with silver nitrate immersion and micro-CT was found more frequently in PFI group. The technique combining resin infiltration and composite restoration might ensure better adhesion prognosis as applied by the sequence of resin infiltration, cavity preparation, and composite filling. This new trial was thought meaningful in minimizing the cavity size and contributing to minimal invasive dentistry.
This study was conducted to evaluate the potential use of milk mineral (MM) as the calcium source for the production of calcium-fortified yogurt. MM was composed of 83% minerals, 7.5% lactose, 3.3% protein, and < 1% fat. Calcium (Ca) content in MM was about 46%; calcium: phosphorous ratio was 1.28:1. The aqueous solubility of Ca increased with the decrease in pH; the solubility at pH 4 and 5 was 98% and 53%, respectively. Ca-fortified yogurt with up to 200 mg Ca/100 mL did not show significant differences in acid production and number of viable cells; however, the viscosity increased significantly (p<0.05) with the increase in Ca levels. Microstructure analysis of Ca-fortified yogurt using confocal scanning laser microscopy indicated that the protein network became denser with increasing fortification with MM. There was no significant difference in the sensory quality between the control and Ca-fortified yogurts. Therefore, MM could be used for the production of Ca-fortified yoghurt without compromising the quality characteristics of yogurt.
Yongwook, Shin;Howon, Park;Juhyun, Lee;Siyoung, Lee
Journal of the korean academy of Pediatric Dentistry
/
v.49
no.2
/
pp.149-157
/
2022
The aim of this study was to evaluate the effects of erythrosine-mediated photodynamic therapy (PDT) on Streptococcus mutans biofilm recovery by counting its colony-forming units (CFUs) and via confocal laser scanning microscopy analysis at different time points following PDT. In PDT, photosensitizer was an erythrosine. S. mutans ATCC25175 biofilms were irradiated using an LED curing light. Chlorhexidine (CHX) was used as positive control. After each antimicrobial treatment, samples were cultured to allow biofilm recovery. Viability was measured by calculating the CFU counts after treatment and after every 3 hours for up to 24 hours. Immediately after treatment, the PDT and CHX groups showed equally significant decreases in S. mutans CFU counts compared to the negative control. After 12 hours of reculture, the PDT group showed no significant difference in the decrease in CFU count compared to the negative control, whereas the CHX group showed significantly lower CFU counts throughout the 24-hour period. Erythrosine-mediated PDT can effectively inhibit S. mutans biofilm formation. However, biofilm recovery occurred earlier in the CHX group after PDT. This study provides insights into the clinical effectiveness of PDT in preventing dental caries.
Gabriela Leite de Souza;Thamara Eduarda Alves Magalhaes;Gabrielle Alves Nunes Freitas;Nelly Xiomara Alvarado Lemus;Gabriella Lopes de Rezende Barbosa;Anielle Christine Almeida Silva;Camilla Christian Gomes Moura
Restorative Dentistry and Endodontics
/
v.47
no.4
/
pp.38.1-38.15
/
2022
Objectives: This study investigated the cytotoxicity, radiopacity, pH, and dentinal tubule penetration of a paste of 1.0% calcium-doped zinc oxide nanocrystals (ZnO:1.0Ca) combined with propylene glycol (PRG) or polyethylene glycol and propylene glycol (PEG-PRG). Materials and Methods: The pastes were prepared by mixing calcium hydroxide [Ca(OH)2] or ZnO:1.0Ca with PRG or a PEG-PRG mixture. The pH was evaluated after 24 and 96 hours of storage in deionized water. Digital radiographs were acquired for radiopacity analysis and bubble counting of each material. The materials were labeled with 0.1% fluorescein and applied to root canals, and images of their dentinal tubule penetration were obtained using confocal laser scanning microscopy. RAW264.7 macrophages were placed in different dilutions of culture media previously exposed to the materials for 24 and 96 hours and tested for cell viability using the MTT assay. Analysis of variance and the Tukey test (α = 0.05) were performed. Results: ZnO:1.0Ca materials showed lower viability at 1:1 and 1:2 dilutions than Ca(OH)2 materials (p < 0.0001). Ca(OH)2 had higher pH values than ZnO:1.0Ca at 24 and 96 hours, regardless of the vehicle (p < 0.05). ZnO:1.0Ca pastes showed higher radiopacity than Ca(OH)2 pastes (p < 0.01). No between-material differences were found in bubble counting (p = 0.0902). The ZnO:1.0Ca pastes had a greater penetration depth than Ca(OH)2 in the apical third (p < 0.0001). Conclusions: ZnO:1.0Ca medicaments presented higher penetrability, cell viability, and radiopacity than Ca(OH)2. Higher values of cell viability and pH were present in Ca(OH)2 than in ZnO:1.0Ca.
Park, Se-Eun;Yi, Kee-Wook;Kim, Hae-Young;Son, Ho-Hyun;Chang, Ju-Hea
Restorative Dentistry and Endodontics
/
v.36
no.4
/
pp.290-299
/
2011
Objectives: The usage of fluoride varnish for a moderate to low caries-risk group has not been well validated. This study aimed to evaluate the preventive and therapeutic efficacies of fluoride varnish on the initiated root caries. Materials and Methods: Ten premolars were sectioned into quarters, further divided into two windows, one of which was painted with Fluor Protector (1,000 ppm fluoride, Ivoclar Vivadent). An initial lesion with a well-preserved surface layer was produced by pH cycling. Scanned line analysis using energy dispersive spectrometry determined the weight percentages of Ca and P in the demineralized layer. Scanning Electron microscopy and confocal laser scanning microscopy (CLSM) evaluated the varnish-applied root surfaces. Results: The mean lesion depth (SD) was 12.3 (2.6) ${\mu}m$ (single cycling) and 19.6 (3.8) ${\mu}m$ (double cycling). Double cycling extended the lesion depth, but induced no more mineral loss than single cycling (p < 0.05). The mean weight percentages of Ca and P between groups with and without varnish were not significantly different (p < 0.05). A CLSM showed varnish remained within 15 ${\mu}m$ of the surface layer. Conclusions: When a mild acid challenge initiated root tissue demineralization, the application of low-concentration fluoride varnish did not influence the lesion depth or the mineral composition of the subsurface lesion.
Water supplied through dental unit waterlines (DUWLs) has been shown to contain high number of bacteria. To reduce the contamination of DUWLs, it is essential to develop effective disinfectants. It is, however, difficulty to obtain proper DUWL samples for studies. The purpose of this study was to establish a simple laboratory model for reproducing DUWL biofilms. The bacteria obtained from DUWLs were cultured in R2A liquid medium for 10 days, and then stored at $-70^{\circ}C$. This stock was inoculated into R2A liquid medium and incubated in batch mode. After 5 days of culturing, it was inoculated into the biofilm formation model developed in this study. Our biofilm formation model comprised of a beaker containing R2A liquid medium and five glass rods attached to DUWL polyurethane tubing. Biofilm was allowed to form on the stir plate and the medium was replaced every 2 days. After 4 days of biofilm formation in the laboratory model, biofilm thickness, morphological characteristics and distribution of the composing bacteria were examined by confocal laser microscopy and scanning electron microscopy. The mean of biofilm accumulation was $4.68{\times}10^4$ colony forming unit/$cm^2$ and its thickness was $10{\sim}14{\mu}m$. In our laboratory model, thick bacterial lumps were observed in some parts of the tubing. To test the suitability of this biofilm model system, the effectiveness of disinfectants such as sodium hypochlorite, hydrogen peroxide, and chlorhexidine, was examined by their application to the biofilm formed in our model. Lower concentrations of disinfectants were less effective in reducing the count of bacteria constituting the biofilm. These results showed that our DUWL biofilm laboratory model was appropriate for comparison of disinfectant effects. Our laboratory model is expected to be useful for various other purposes in further studies.
The water discharged from dental unit waterlines (DUWLs) is heavily contaminated with bacteria. The development of efficient disinfectants is required to maintain good quality DUWL water. The purpose of this study was to establish a DUWL biofilm model using well-plates to confirm the effectiveness of disinfectants in the laboratory. Bacteria were obtained from the water discharged from DUWLs and incubated in R2A liquid medium for 10 days. The bacterial solution cultured for 10 days was made into stock and these stocks were incubated in R2A broth and batch mode for 5 days. Batch-cultured bacterial culture solution and polyurethane tubing sections were incubated in 12-well plates for 4 days. Biofilm accumulation was confirmed through plating on R2A solid medium. In addition, the thickness of the biofilm and the shape and distribution of the constituent bacteria were confirmed using confocal laser microscopy and scanning electron microscopy. The average accumulation of the cultured biofilm over 4 days amounted to $1.15{\times}10^7CFU/cm^2$. The biofilm was widely distributed on the inner surface of the polyurethane tubing and consisted of cocci, short-length rods and medium-length rods. The biofilm thickness ranged from $2{\mu}m$ to $7{\mu}m$. The DUWL biofilm model produced in this study can be used to develop disinfectants and study DUWL biofilm-forming bacteria.
The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, $H_2SO_4$ and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven ($37^{\circ}C$). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of $H_2SO_4$ and temperature of acid-solution. Acid-treatment conditions (concentration of $H_2SO_4$, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with $90^{\circ}C$ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.
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