Browsing by Author "Arthur, Rodrigo A."

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    Biodegradable nanofibrous drug delivery systems: effects of metronidazole and ciprofloxacin on periodontopathogens and commensal oral bacteria
    (Springer-Verlag, 2014-12) Bottino, Marco C.; Arthur, Rodrigo A.; Waeiss, R. Aaron; Kamocki, Krzysztof; Gregson, Karen S.; Gregory, Richard L.; Department of Restorative Dentistry, IU School of Dentistry
    OBJECTIVES: The purposes of this study were to fabricate biodegradable polydioxanone (PDS II®) electrospun periodontal drug delivery systems (hereafter referred to as matrices) containing either metronidazole (MET) or ciprofloxacin (CIP) and to investigate the effects of antibiotic incorporation on both periodontopathogens and commensal oral bacteria. MATERIALS AND METHODS: Fibrous matrices were processed from PDS polymer solution by electrospinning. Antibiotic-containing PDS solutions were prepared to obtain four distinct groups: 5 wt.% MET, 25 wt.% MET, 5 wt.% CIP, and 25 wt.% CIP. Pure PDS was used as a control. High-performance liquid chromatography (HPLC) was done to evaluate MET and CIP release. Dual-species biofilms formed by Lactobacillus casei (Lc) and Streptococcus salivarius (Ss) were grown on the surface of all electrospun matrices. After 4 days of biofilm growth, the viability of bacteria on biofilms was assessed. Additionally, antimicrobial properties were evaluated against periodontopathogens Fusobacterium nucleatum (Fn) and Aggregatibacter actinomycetemcomitans (Aa) using agar diffusion assay. RESULTS: A three-dimensional interconnected porous network was observed in the different fabricated matrices. Pure PDS showed the highest fiber diameter mean (1,158 ± 402 nm) followed in a descending order by groups 5 wt.% MET (1,108 ± 383 nm), 25 wt.% MET (944 ± 392 nm), 5 wt.% CIP (871 ± 309 nm), and 25 wt.% CIP (765 ± 288 nm). HPLC demonstrated that groups containing higher amounts (25 wt.%) of incorporated drugs released more over time, while those with lower levels (5 wt.%) the least. No inhibitory effect of the tested antibiotics was detected on biofilm formation by the tested commensal oral bacteria. Meanwhile, CIP-containing matrices inhibited growth of Fn and Aa. CONCLUSION: CIP-containing matrices led to a significant inhibition of periodontopathogens without negatively impairing the growth of periodontal beneficial bacteria. CLINICAL RELEVANCE: Based on the proven in vitro inhibition of periodontitis-related bacteria, future in vivo research using relevant animal models is needed to confirm the effectiveness of these drug delivery systems.
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    Farnesol Delivery via Polymeric Nanoparticle Carriers Inhibits Cariogenic Cross-Kingdom Biofilms and Prevents Enamel Demineralization
    (Wiley, 2022) Ito, Tatsuro; Sims, Kenneth R., Jr.; Liu, Yuan; Xiang, Zhenting; Arthur, Rodrigo A.; Hara, Anderson T.; Koo, Hyun; Benoit, Danielle S. W.; Klein, Marlise I.; Biomedical and Applied Sciences, School of Dentistry
    Streptococcus mutans and Candida albicans are frequently detected together in the plaque from patients with early childhood caries (ECC) and synergistically interact to form a cariogenic cross-kingdom biofilm. However, this biofilm is difficult to control. Thus, to achieve maximal efficacy within the complex biofilm microenvironment, nanoparticle carriers have shown increased interest in treating oral biofilms in recent years. Here, we assessed the anti-biofilm efficacy of farnesol (Far), a hydrophobic antibacterial drug and repressor of Candida filamentous forms, against cross-kingdom biofilms employing drug delivery via polymeric nanoparticle carriers (NPCs). We also evaluated the effect of the strategy on teeth enamel demineralization. The farnesol-loaded NPCs (NPC+Far) resulted in a 2-log CFU/mL reduction of S. mutans and C. albicans (hydroxyapatite disc biofilm model). High-resolution confocal images further confirmed a significant reduction in exopolysaccharides, smaller microcolonies of S. mutans, and no hyphal form of C. albicans after treatment with NPC+Far on human tooth enamel (HT) slabs, altering the biofilm 3D structure. Furthermore, NPC+Far treatment was highly effective in preventing enamel demineralization on HT, reducing lesion depth (79% reduction) and mineral loss (85% reduction) versus vehicle PBS-treated HT, while NPC or Far alone had no differences with the PBS. The drug delivery via polymeric NPCs has the potential for targeting bacterial-fungal biofilms associated with a prevalent and costly pediatric oral disease, such as ECC.
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    Objective and Quantitative Assessment of Caries Lesion Activity
    (Elsevier, 2018) Ando, Masatoshi; Fontana, Margherita; Eckert, George J.; Arthur, Rodrigo A.; Zhang, Hui; Zero, Domenick T.; Cariology, Operative Dentistry and Dental Public Health, School of Dentistry
    Objectives Evaluate the ability of objectively measured specular reflection, roughness, and fluorescence change during dehydration to assess caries lesion activity. Methods One hundred ninety-five ground/polished 3 × 3 × 2 mm sound human enamel specimens were divided into three groups and demineralized using a multispecies microbial caries model for 3, 6, or 9 days; and then remineralized with 1100 ppm-F as NaF solution for 10 days using a pH-cyclic model. Reflection (amplitude: %), roughness (Ra: μm), fluorescence change during dehydration (ΔQ: %×mm2), and microfocus computed tomography [μ-CT: lesion volume (μm3)] were measured for sound, demineralized and remineralized enamel. The surface was hydrated and fluorescence images were acquired at 1 s intervals for 10 s (ΔQ10). During image acquisition, surface was dehydrated with continuous compressed air. Changes-in-ΔQ per second (ΔQD: %×mm2/sec) at 5 (ΔQD5) and 10 s (ΔQD10) were obtained. Results Reflection decreased from sound to demineralized groups (p < 0.0001); remineralized groups were higher than demineralized groups (p < 0.001), but not different from sound (p > 0.32). Roughness increased from sound to demineralized groups (p < 0.0001) and remineralized groups were also higher than sound (p < 0.0001). ΔQ10, ΔQD5 and ΔQD10 increased from sound to demineralized groups (p < 0.0001), and remineralized groups decreased compared to demineralized groups (p < 0.05), but was higher than sound (p < 0.0001). The correlations of μ-CT with reflection, roughness, and ΔQ10 were −0.63, 0.71, and 0.82, respectively (p < 0.0001). Conclusions Reflection, roughness and ΔQ could distinguish between sound and demineralized enamel. Reflection and ΔQ were able to distinguish between demineralized and remineralized enamel. Clinical significance Determination of caries activity, whether a lesion is active or inactive, is an essential and critical component of caries diagnosis. However, especially for enamel lesions, it is difficult to estimate without longitudinal follow-up. Reflection, roughness and fluorescence change during dehydration have the potential to measure caries lesion activity at a-single-appointment.
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    Salivary epithelial cells as model to study immune response against cutaneous pathogens
    (Wiley, 2014-02) Negrini, Thais C.; Arthur, Rodrigo A.; Waeiss, Robert A.; Carlosa, Iracilda Z.; Srinivasan, Mythily; Oral Pathology, Medicine and Radiology, School of Dentistry
    The human skin not only provides passive protection as a physical barrier against external injury, but also mediates active surveillance via epidermal cell surface receptors that recognize and respond to potential invaders. Primary keratinocytes and immortalized cell lines, the commonly used sources to investigate immune responses of cutaneous epithelium are often difficult to obtain and/or potentially exhibit changes in cellular genetic make-up. Here we investigated the possibility of using salivary epithelial cells (SEC) to evaluate the host response to cutaneous microbes. Elevated secretion of IFN-γ and IL-12 was observed in the SEC stimulated with Staphylococcus aureus, a transient pathogen of the skin, as mono species biofilm as compared to SEC stimulated with a commensal microbe, the Staphylococcus epidermidis. Co-culture of the SEC with both microbes as dual species biofilm elicited maximum cytokine response. Stimulation with S. aureus alone but not with S. epidermidis alone induced maximum toll-like receptor-2 (TLR-2) expression in the SEC. Exposure to dual species biofilm induced a sustained upregulation of TLR-2 in the SEC for up to an hour. The data support novel application of the SEC as efficient biospecimen that may be used to investigate personalized response to cutaneous microflora.
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    Spatial mapping of polymicrobial communities reveals a precise biogeography associated with human dental caries
    (National Academy of Sciences, 2020-06-02) Kim, Dongyeop; Barraza, Juan P.; Arthur, Rodrigo A.; Hara, Anderson; Lewis, Karl; Liu, Yuan; Scisci, Elizabeth L.; Hajishengallis, Evlambia; Whiteley, Marvin; Koo, Hyun; Anatomy and Cell Biology, School of Medicine
    Tooth decay (dental caries) is a widespread human disease caused by microbial biofilms. Streptococcus mutans, a biofilm-former, has been consistently associated with severe childhood caries; however, how this bacterium is spatially organized with other microorganisms in the oral cavity to promote disease remains unknown. Using intact biofilms formed on teeth of toddlers affected by caries, we discovered a unique 3D rotund-shaped architecture composed of multiple species precisely arranged in a corona-like structure with an inner core of S. mutans encompassed by outer layers of other bacteria. This architecture creates localized regions of acidic pH and acute enamel demineralization (caries) in a mixed-species biofilm model on human teeth, suggesting this highly ordered community as the causative agent. Notably, the construction of this architecture was found to be an active process initiated by production of an extracellular scaffold by S. mutans that assembles the corona cell arrangement, encapsulating the pathogen core. In addition, this spatial patterning creates a protective barrier against antimicrobials while increasing bacterial acid fitness associated with the disease-causing state. Our data reveal a precise biogeography in a polymicrobial community associated with human caries that can modulate the pathogen positioning and virulence potential in situ, indicating that micron-scale spatial structure of the microbiome may mediate the function and outcome of host–pathogen interactions.