Potentials of Mouthwashes in Disinfecting Cariogenic Bacteria and Biofilms Leading to Inhibition of Caries

Objectives:The aim of this study was to compare the effects of certain commercially available mouthwashes on cariogenic bacteria and biofilms, following the acquisition of inhibition potentials of caries.Materials and Methods:Mouthwashes containing I) chlorhexidine gluconate (CHG; 0.0005% w/v), II) benzethonium chloride (BTC; 0.01% w/v), III) an essential oil (Listerine), and IV) povidone-iodine (PVP-I; 0.035% w/v) were tested on planktonic cariogenic bacteria, biofilms, and an ex vivo caries model. Bacterial aliquots were inoculated with each solution separately and vortexed for 10 seconds at room temperature. Bacterial viability was subsequently investigated by fluorescence microscopy (FM) after staining with a BacLight viability kit and the number of colony-forming units (CFUs) was counted. Similarly, mouthwash solutions were applied to artificial cariogenic biofilms, and bacterial viability of the biofilms was investigated as stated above. Inhibition potentials of two selected mouthwashes of carious lesions were investigated using biofilm-induced caries and a secondary caries model. In all steps, a phosphate-buffered saline (PBS) solution was included as a control.Results:Planktonic cariogenic bacteria and bacteria embedded in biofilms were killed in remarkably large numbers with Listerine and PVP-I treatment compared to PBS and other gargles. CFU counts also showed significant reduction after treatment with Listerine and PVP-I compared to other solutions (P<0.05). Listerine also displayed significant (P<0.05) inhibition effects in preventing the progression of demineralization.Conclusion:Bactericidal potencies of the mouthwashes varied significantly, suggesting that mouthwashes like Listerine can be useful for the prevention of caries and secondary caries.

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Novel Nanocomposite Inhibiting Caries at the Enamel Restoration Margins in an In Vitro Saliva-Derived Biofilm Secondary Caries Model

International Journal of Molecular Sciences ◽

10.3390/ijms21176369 ◽

2020 ◽

Vol 21 (17) ◽

pp. 6369

Author(s):

Wen Zhou ◽

Xinyu Peng ◽

Xuedong Zhou ◽

Andrea Bonavente ◽

Michael D. Weir ◽

...

Keyword(s):

Low Ph ◽

The Novel ◽

Antibacterial Effects ◽

Ion Release ◽

Secondary Caries ◽

Colony Forming Units ◽

Caries Model ◽

Bovine Enamel ◽

First Time

Secondary caries often occurs at the tooth-composite margins. This study developed a novel bioactive composite containing DMAHDM (dimethylaminohexadecyl methacrylate) and NACP (nanoparticles of amorphous calcium phosphate), inhibiting caries at the enamel restoration margins in an in vitro saliva-derived biofilm secondary caries model for the first time. Four composites were tested: (1) Heliomolar nanocomposite, (2) 0% DMAHDM + 0% NACP, (3) 3% DMAHDM + 0% NACP, (D) 3% DMAHDM + 30% NACP. Saliva-derived biofilms were tested for antibacterial effects of the composites. Bovine enamel restorations were cultured with biofilms, Ca and P ion release of nanocomposite and enamel hardness at the enamel restoration margins was measured. Incorporation of DMAHDM and NACP into composite did not affect the mechanical properties (p > 0.05). The biofilms’ CFU (colony-forming units) were reduced by 2 logs via DMAHDM (p < 0.05). Ca and P ion release of the nanocomposite was increased at cariogenic low pH. Enamel hardness at the margins for DMAHDM group was 25% higher than control (p < 0.05). With DMAHDM + NACP, the enamel hardness was the greatest and about 50% higher than control (p < 0.05). Therefore, the novel composite containing DMAHDM and NACP was strongly antibacterial and inhibited enamel demineralization, resulting in enamel hardness at the margins under biofilms that approached the hardness of healthy enamel.

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Silica Nanoparticles to Polish Tooth Surfaces for Caries Prevention

Journal of Dental Research ◽

10.1177/154405910808701007 ◽

2008 ◽

Vol 87 (10) ◽

pp. 980-983 ◽

Cited By ~ 30

Author(s):

R.M. Gaikwad ◽

I. Sokolov

Keyword(s):

Silica Nanoparticles ◽

Ex Vivo ◽

Bacterial Attachment ◽

Nanometer Scale ◽

Human Teeth ◽

Cariogenic Bacteria ◽

Force Microscopy ◽

Atomic Force ◽

Tooth Surfaces ◽

Scale Roughness

Although silica particles have been used for tooth polishing, polishing with nanosized particles has not been reported. Here we hypothesize that such polishing may protect tooth surfaces against the damage caused by cariogenic bacteria, because the bacteria can be easily removed from such polished surfaces. This was tested on human teeth ex vivo. The roughness of the polished surfaces was measured with atomic force microscopy (AFM). A considerably lower nanometer-scale roughness was obtained when silica nanoparticles were used to polish the tooth surfaces, as compared with conventional polishing pastes. Bacterial attachment to the dental surfaces was studied for Streptococcus mutans, the most abundant cariogenic bacteria. We demonstrated that it is easier to remove bacteria from areas polished with silica nanoparticles. The results demonstrate the advantage of using silica nanoparticles as abrasives for tooth polishing.

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Ex Vivo-Generated CD36+ Erythroid Progenitors Are Highly Permissive to Human Parvovirus B19 Replication

Journal of Virology ◽

10.1128/jvi.02247-07 ◽

2007 ◽

Vol 82 (5) ◽

pp. 2470-2476 ◽

Cited By ~ 69

Author(s):

Susan Wong ◽

Ning Zhi ◽

Claudia Filippone ◽

Keyvan Keyvanfar ◽

Sachiko Kajigaya ◽

...

Keyword(s):

Progenitor Cells ◽

Real Time ◽

Cell Lines ◽

Ex Vivo ◽

Parvovirus B19 ◽

Erythroid Progenitor ◽

Hematopoietic Stem ◽

Human Virus ◽

Erythroid Progenitor Cells ◽

Large Numbers

ABSTRACT The pathogenic parvovirus B19 (B19V) has an extreme tropism for human erythroid progenitor cells. In vitro, only a few erythroid leukemic cell lines (JK-1 and KU812Ep6) or megakaryoblastoid cell lines (UT7/Epo and UT7/Epo-S1) with erythroid characteristics support B19V replication, but these cells are only semipermissive. By using recent advances in generating large numbers of human erythroid progenitor cells (EPCs) ex vivo from hematopoietic stem cells (HSCs), we produced a pure population of CD36+ EPCs expanded and differentiated from CD34+ HSCs and assessed the CD36+ EPCs for their permissiveness to B19V infection. Over more than 3 weeks, cells grown in serum-free medium expanded more than 800,000-fold, and 87 to 96% of the CD36+ EPCs were positive for globoside, the cellular receptor for B19V. Immunofluorescence (IF) staining showed that about 77% of the CD36+ EPCs were positive for B19V infection, while about 9% of UT7/Epo-S1 cells were B19V positive. Viral DNA detected by real-time PCR increased by more than 3 logs in CD36+ EPCs; the increase was 1 log in UT7/Epo-S1 cells. Due to the extensive permissivity of CD36+ EPCs, we significantly improved the sensitivity of detection of infectious B19V by real-time reverse transcription-PCR and IF staining 100- and 1,000-fold, respectively, which is greater than the sensitivity of UT7/Epo-S1 cell-based methods. This is the first description of an ex vivo method to produce large numbers of EPCs that are highly permissive to B19V infection and replication, offering a cellular system that mimics in vivo infection with this pathogenic human virus.

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In Vitro Effect of Local Anesthetics onCandida albicansGerm Tube Formation

Infectious Diseases in Obstetrics and Gynecology ◽

10.1155/s1064744994000074 ◽

1994 ◽

Vol 1 (4) ◽

pp. 193-197 ◽

Cited By ~ 5

Author(s):

Acácio Rodrigues ◽

Cidália Pina Vaz ◽

A. Freitas Fonseca ◽

J. Martinez de Oliveira ◽

Henrique Barros

Keyword(s):

Germ Tube ◽

Inhibitory Effect ◽

Tube Formation ◽

Vaginal Candidiasis ◽

Sabouraud Agar ◽

Colony Forming Units ◽

Vitro Effect ◽

Phosphate Buffered Saline ◽

Dose Dependent

Objective:This study was planned to clarify the in vitro effect of lidocaine and bupivacaine on germ tube formation byCandida albicansisolates from cases of clinical vaginal candidiasis.Methods:FourteenC. albicansstrains (clinical vaginal isolates) were grown on Sabouraud agar for 24 h at 37℃ and tested as follows: 100 μl of a yeast suspension [105colony forming units (CFU)/ml of phosphate buffered saline (PBS)] was added to 500 μl of fresh human serum with lidocaine or bupivacaine (pure salts) in serial concentrations. The test was run in duplicate. Controls were prepared for each strain. After 4 h of incubation at 37℃, samples were taken from each vial and 200 yeasts were counted in a counting chamber. The pH of each suspension was measured.Results:The results are given as the mean of the 2 readings and are expressed as the percentage of blastoconidia with germ tubes/total blastoconidia.Conclusions:Our experiments show that both lidocaine and bupivacaine have a dose-dependent inhibitory effect, pH-independent, on germ tube formation byC. albicansand that both drugs seem to be promising in the treatment of genital candidiasis due to the combination of anesthetic and antifungal properties.

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Acute- and chronic-phase chronic myelogenous leukemia colony-forming units are highly sensitive to the growth inhibitory effects of c-myb antisense oligodeoxynucleotides

Blood ◽

10.1182/blood.v79.8.1956.1956 ◽

1992 ◽

Vol 79 (8) ◽

pp. 1956-1961 ◽

Cited By ~ 72

Author(s):

MZ Ratajczak ◽

N Hijiya ◽

L Catani ◽

K DeRiel ◽

SM Luger ◽

...

Keyword(s):

Chronic Myelogenous Leukemia ◽

Colony Formation ◽

Ex Vivo ◽

Blast Crisis ◽

Chronic Phase ◽

Myelogenous Leukemia ◽

Antisense Oligodeoxynucleotides ◽

Rt Pcr ◽

Colony Forming Units

Abstract We have previously demonstrated that malignant hematopoietic colony- forming units (CFUs) may be purged from normal CFU by exposure to c-myb antisense oligodeoxynucleotides (oligomers). This novel strategy appeared particularly promising for patients with chronic myelogenous leukemia (CML) in blast crisis, since in some cases complete elimination of bcr-abl-expressing cells was accomplished. We have examined 11 additional patients, including seven in chronic phase, in order to extend these initial observations. We sought in particular to determine if elimination of bcr-abl-expressing clones was a usual event. Exposure of CML cells to c-myb antisense oligomers resulted in inhibition of CFU-granulocyte, macrophage (CFU-GM)-derived colony formation in eight of 11 (73%) cases evaluated. Inhibition was antisense sequence-specific, dose-dependent, ranged between 58% and 93%, and was statistically significant (P less than or equal to .03) in seven of the eight cases. In two cases, CFU-granulocyte, erythrocyte, monocyte, megakaryocyte (CFU-GEMM)-derived colony formation was also examined and found to be inhibited by the c-myb antisense oligomers in a sequence-specific manner. To determine whether CML CFU had been reduced or eliminated after exposure to the antisense oligomers, we examined cells in the residual colonies for bcr-abl mRNA expression using a reverse transcription-polymerase chain reaction detection technique (RT-PCR). Eight cases were evaluated and in each case where antisense myb inhibited growth, bcr-abl expression as detected by RT- PCR was either greatly decreased or nondetectable. No residual leukemic CFU were demonstrable on replating of treated cells. These results suggest that c-myb antisense oligomers substantially inhibit the growth and survival of CML CFU in both chronic and blast phase of disease. They may therefore prove useful for both ex vivo and in vivo treatment of CML.

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EFFECTS OF N-ACETYL-CYSTEINE SUPPLEMENTATION ON EX-VIVO CLONOGENICITY AND OXIDATIVE PROFILE OF LINEAGE-COMMITTED HEMATOPOIETIC STEM/PROGENITOR CELLS

Jurnal Teknologi ◽

10.11113/jt.v80.11419 ◽

2018 ◽

Vol 80 (3) ◽

Cited By ~ 1

Author(s):

Chan Chin Yi ◽

Zariyantey Abd Hamid ◽

Izatus Shima Taib ◽

Tan Hui Yee ◽

Muhd Khairul Akmal Wak Harto ◽

...

Keyword(s):

Progenitor Cells ◽

Ex Vivo ◽

Oxidative Status ◽

Oxygen Species ◽

Hematopoietic Stem ◽

Colony Forming Units ◽

Stem And Progenitor Cells ◽

Acetyl Cysteine ◽

Antioxidant Supplement ◽

B Lymphoid

Hematopoietic stem and progenitor cells (HSPCs) are exposed to oxidative damage acquired during ex vivo expansion which affects their therapeutic potency. Efforts to overcome this limitation includes the use of antioxidants. The effects of N-Acetyl-Cysteine (NAC) supplementation for 48 hours on maintenance of ex vivo HSPCs was investigated by examining the cell viability at concentrations of 0.125 µM, 0.25 µM, 0.5 µM, 1.0 µM and 2.0 µM, followed by clonogenicity and oxidative status assessments of lineage-committed progenitors (myeloid, erythroid and pre-B lymphoid) at selected NAC concentrations (0.25 µM, 0.5 µM, 2.0 µM). NAC supplementation significantly (p< 0.05) enhanced viability of HSPC at 0.25 µM, 0.5 µM, 2.0 µM. The clonogenicity of each progenitor was not affected as no significant changes of Colony Forming Units (CFUs) counts was noted between NAC-supplemented group than control. NAC showed no significant effects on reactive oxygen species (ROS), glutathione (GSH) and superoxide dismutase (SOD) levels of respective progenitors as compared to control. Conclusively, NAC shows potential property as antioxidant supplement for ex vivo maintenance of HSPCs by promoting survivability and maintaining clonogenicity.

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Collection of pluripotential hematopoietic stem cells by cytapheresis

Blood ◽

10.1182/blood.v59.4.822.822 ◽

1982 ◽

Vol 59 (4) ◽

pp. 822-827 ◽

Cited By ~ 39

Author(s):

LC Lasky ◽

RC Ash ◽

JH Kersey ◽

ED Zanjani ◽

J McCullough

Keyword(s):

Stem Cells ◽

Collection Efficiency ◽

Hematopoietic Stem ◽

Hematopoietic Reconstitution ◽

Large Numbers ◽

Colony Forming Units ◽

Peripheral Stem Cells ◽

And Storage ◽

Marrow Ablation ◽

Cell Colony

Abstract Successful complete hematopoietic reconstitution (CHR) using nonleukemic peripheral stem cells (PSC) after marrow ablation has been reported in animals but not man. Previous studies of cytapheresis products from humans, as a prelude to use for CHR, have documented the presence of committed myeloid (CFU-GM) and erythroid (BFU-E) precursors. We have examined mononuclear cell (MNC) products collected on the Fenwal CS3000 Blood Cell Separator for these plus the more primitive mixed (granulo-, erythro-, mono-, and megakaryocytic) cell colony-forming units (CFU-GEMM) and for various lymphocytic subpopulations (LSP). One to two-hour products contained 36 +/- 7 CFU- GEMM/10(6) MNC (mean +/- SE, n = 8) or 490 +/- 131/ml product. This compared favorably with blood (23 +/- .4/10(6) MNC or 46 +/- 8/ml, n = 14) and bone marrow (146 +/- 58/10(6) MNC, n = 12). Collection efficiency for E-rosette-positive cells approximated that for total lymphocytes and was variable for other LSP. Recovery of CFU-GEMM after freezing in 10% dimethylsulfoxide at a controlled rate and storage in liquid N2 was 54% +/- 8% (n = 8). Cytapheresis collection of large numbers of pluripotent hematopoietic precursors and demonstration of adequate recovery of these after cryopreservation, both previously unreported, are significant steps toward eventual CHR using nonleukemic PSC.

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Ex-Vivo Expansion of Multipotent CD133+ Stem Cells with VEGF, FLT3l and SCGF.

Blood ◽

10.1182/blood.v104.11.4147.4147 ◽

2004 ◽

Vol 104 (11) ◽

pp. 4147-4147

Author(s):

Sonja Loges ◽

Martin Butzal ◽

Uta Fischer ◽

Ursula M. Gehling ◽

Dieter K. Hossfeld ◽

...

Keyword(s):

Stem Cells ◽

Endothelial Cells ◽

Stem Cell ◽

Culture System ◽

Ex Vivo ◽

Ex Vivo Expansion ◽

Facs Analysis ◽

Hematopoietic Stem ◽

Large Numbers

Abstract The rare CD133+ stem cell population contains both hematopoietic and endothelial progenitors. Successful ex-vivo expansion of this multipotent population would therefore be of great benefit in many clinical settings including stem cell transplantation and gene therapy. We developed a cell culture system containing the recombinant human cytokines vascular endothelial growth factor (VEGF), FLT3 ligand (FLT3L) and stem cell growth factor (SCGF) for ex-vivo expansion of purified human CD133+ stem cells obtained from leukapheresis products from patients pre-treated with G-CSF. FACS analysis, colony assays and NOD-SCID transplantation studies were performed to monitor stem cell and endothelial phenotype in-vitro and in-vivo. Cultivation with VEGF, FLT3L and SCGF induced a mean 2200-fold increase of total cell counts in 5 weeks. FACS analysis revealed persistence of 6–15% CD133+ stem cells indicating proliferation and survival of primitive hematopoietic stem cells. 5–6% of the proliferating cells expressed the endothelial markers CD144 (VE-Cadherin) and von-Willebrand factor (vWF). Ex-vivo expanded stem cells could be differentiated into adherent endothelial cells after withdrawal of SCGF and FLT3L allowing generation of large numbers of endothelial cells. Colony-assays showed an increase of hematopoietic and endothelial colonies after 5 weeks of ex-vivo expansion indicating simultaneous proliferation of hematopoietic and endothelial precursors under the established culture conditions (CFU-E 60-fold, CFU-GEMM 48-fold, CFU-GM 59-fold, CFU-G 99-fold, CFU-M 1356-fold and CFU-EC 1843-fold). To assess in-vivo functionality, hematopoietic stem cells expanded ex-vivo for 7, 14, 21 and 32 days were transplanted into sublethally irradiated NOD-SCID mice. For each expansion period, the mean percentage of anti-human CD45 positive bone marrow cells 3 months post-transplantation was 11, 3, 3 and 1%, respectively. Human CD45+ cells for each set of experiments contained a mean of 15, 26, 8 and 32% T-cells (CD3+), 9, 0, 7 and 21% B-cells (CD19+), 24, 2, 2 and 11% monocytes (CD14+), 21, 3, 1 and 12% granulocytes (CD33+) and 19, 37, 44 and 24% stem cells (CD34+) (d7 (n=5), d14 (n=4), d21 (n=7) and d32 (n=6) respectively). Our experiments showed multilineage engraftment of human stem cells expanded for more than 4 weeks ex-vivo. Therefore our culture system provides a tool to generate large numbers of human stem and endothelial cells for clinical purposes.

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The Antibacterial Effect of Ethanol Extract of Polish Propolis on Mutans Streptococci and Lactobacilli Isolated from Saliva

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/681891 ◽

2013 ◽

Vol 2013 ◽

pp. 1-12 ◽

Cited By ~ 21

Author(s):

Arkadiusz Dziedzic ◽

Robert Kubina ◽

Robert D. Wojtyczka ◽

Agata Kabała-Dzik ◽

Marta Tanasiewicz ◽

...

Keyword(s):

Ex Vivo ◽

Antibacterial Properties ◽

Ethanol Extract ◽

Mutans Streptococci ◽

Diffusion Method ◽

Extracellular Polysaccharides ◽

Cariogenic Bacteria ◽

Slide Test ◽

The Mean ◽

Alamarblue Assay

Dental caries occurrence is caused by the colonization of oral microorganisms and accumulation of extracellular polysaccharides synthesized byStreptococcus mutanswith the synergistic influence ofLactobacillusspp. bacteria. The aim of this study was to determineex vivothe antibacterial properties of ethanol extract of propolis (EEP), collected in Poland, against the main cariogenic bacteria: salivary mutans streptococci and lactobacilli. The isolation of mutans streptococci group bacteria (MS) andLactobacillusspp. (LB) from stimulated saliva was performed by in-office CRT bacteria dip slide test. The broth diffusion method and AlamarBlue assay were used to evaluate the antimicrobial activity of EEP, with the estimation of its minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The biochemical composition of propolis components was assessed. The mean MIC and MBC values of EEP, in concentrations ranging from 25 mg/mL to 0.025 mg/mL, for the MS and LB were found to be 1.10 mg/mLversus0.7 mg/mL and 9.01 mg/mLversus5.91 mg/mL, respectively. The exposure to an extract of Polish propolis affected mutans streptococci andLactobacillusspp. viability, exhibiting an antibacterial efficacy on mutans streptococci group bacteria and lactobacilli saliva residents, while lactobacilli were more susceptible to EEP. Antibacterial measures containing propolis could be the local agents acting against cariogenic bacteria.

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