The oral microbiome is a diverse ecosystem that plays a critical role in health and disease and contains numerous bacterial species capable of metabolizing host-derived glycans, particularly sialic acids. Sialidase enzymes can be produced by both commensal and pathogenic bacteria, influencing biofilm formation and host-interactions. To investigate how sialidase activity might influence the oral microbiome we conducted a series of in vitro polymicrobial biofilm experiments and assessed community composition using 16S rRNA sequencing. As a first step, we tested modified Oxford Nanopore Technology (ONT) primers using an in-house sequencing workflow and compared it to the standard Illumina MiSeq primers. Through in silico and in vitro assessments, we identified primer bias in the standard ONT 16S primers and designed human oral microbiome (HOM) modified primers (HOM_27F-YM/1492R-D) to improve taxonomic resolution, achieving results comparable to the gold-standard Illumina 16S primers, particularly for key oral genera. These HOM-optimized primers had an overall lower error rate (3.4%) and generated community profiles that closely matched to those produced by Illumina. We then used the same ONT workflow and modified 16S primers to evaluate the effect of sialidase inhibitors Oseltamivir and 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (DANA) on Hydroxyapatite (HA)-coated minimum biofilm eradication concentration (MBEC) assay plate derived plaque biofilms from a whole plaque community model. Inhibitor-treated biofilms exhibited differences in relative abundance depending on the inhibitor combination used, with increased abundance of Streptococcus with Oseltamivir alone and Fusobacterium with both inhibitors combined (Kruskal-Wallis cutoff = 0.05, LDA > 2). These findings demonstrate that ONT-based 16S sequencing with HOM-modified primers suggested that sialidase activity can modulate microbial community structure in plaque biofilms.

The oral microbiome is a diverse ecosystem that plays a critical role in health and disease and contains numerous bacterial species capable of metabolizing host-derived glycans, particularly sialic acids. Sialidase enzymes can be produced by both commensal and pathogenic bacteria, influencing biofilm formation and host-interactions. To investigate how sialidase activity might influence the oral microbiome we conducted a series of in vitro polymicrobial biofilm experiments and assessed community composition using 16S rRNA sequencing. As a first step, we optimized and validated Oxford Nanopore Technology (ONT) as a sequencing platform using an in-house sequencing workflow and compared it to the standard Illumina MiSeq technology. Through in silico and in vitro assessments, we identified primer bias in the standard ONT 16S primers and designed human oral microbiome (HOM) modified primers (HOM_27F-YM/1492R-D) to improve taxonomic resolution, achieving results comparable to the gold-standard Illumina 16S primers, particularly for key oral taxa. These HOM-optimized primers had an overall lower error rate and generated community profiles that closely matched to those produced by Illumina. We then used the same ONT workflow and modified 16S primers to evaluate the effect of sialidase inhibitors Oseltamivir and 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (DANA) on Hydroxyapatite (HA)-coated minimum biofilm eradication concentration (MBEC) assay plate derived plaque biofilms from a whole plaque community model. Inhibitor-treated biofilms exhibited differences in relative abundance depending on the inhibitor combination used, with increased abundance of Streptococcus with Oseltamivir alone and Fusobacterium with both inhibitors combined. These findings demonstrate that ONT-based 16S sequencing is a viable, scalable method for oral microbiome research and suggest that sialidase activity can modulate microbial community structure in plaque biofilms.

Background: The aim of this study was to investigate and visualize the anti-inflammatory and anti-bacterial effects of different oral care products using an infected and inflamed 3D tissue-engineered gingival mucosal model. Methods: A 3D full-thickness oral mucosal model was engineered inside tissue culture inserts using collagen hydrogels populated with human gingival fibroblasts and THP-1 monocytes and layered with oral epithelial cell lines. Oral saliva bacteria were cultured and added to the surface of the models and inflammation was further simulated with lipopolysaccharide (LPS) of Escherichia coli. The 3D models were exposed to three different types of toothpastes, a chlorhexidine antiseptic mouthwash, different antibiotics, and a mechanical rinse with phosphate-buffered saline (PBS) prior to biological evaluation using the PrestoBlue tissue viability assay, histology, optical coherence tomography (OCT), confocal microscopy, and measurement of the release of the inflammatory markers IL-1β, IL-6, and IL-8 with ELISA. Results: Multiple-endpoint analyses of the infected oral mucosal models treated with different anti-bacterial agents showed consistent outcomes in terms of tissue viability, histology, OCT, and confocal microscopy findings. In terms of anti-inflammatory testings, the positive control group showed the highest level of inflammation compared with all other groups. Depending on the anti-bacterial and anti-inflammatory potential of the test groups, different levels of inflammation were observed in the test groups. Conclusions: The inflamed 3D oral mucosal model developed in this study has the potential to be used as a suitable in vitro model for testing the biocompatibility, anti-inflammatory, and anti-bacterial properties of oral care products including mouthwashes and toothpastes. The results of this study indicate that the chlorhexidine mouthwash has both anti-bacterial and cytotoxic effects on the 3D oral mucosal model. Hyaluronic-acid-containing toothpaste has significant anti-bacterial and anti-inflammatory effects on the 3D oral mucosal model.

As one of the most prevalent infective diseases worldwide, it is crucial that we not only know the constituents of the oral microbiome in dental caries but also understand its functionality. Herein, we present a reproducible meta‐analysis to effectively report the key components and the associated functional signature of the oral microbiome in dental caries. Publicly available sequencing data were downloaded from online repositories and subjected to a standardized analysis pipeline before analysis. Meta‐analyses identified significant differences in alpha and beta diversities of carious microbiomes when compared to healthy ones. Additionally, machine learning and receiver operator characteristic analysis showed an ability to discriminate between healthy and disease microbiomes. We identified from importance values, as derived from random forest analyses, a group of genera, notably containing Selenomonas, Aggregatibacter, Actinomyces and Treponema, which can be predictive of dental caries. Finally, we propose the most appropriate study design for investigating the microbiome of dental caries by synthesizing the studies, which had the most accurate differentiation based on random forest modelling. In conclusion, we have developed a non‐biased, reproducible pipeline, which can be applied to microbiome meta‐analyses of multiple diseases, but importantly we have derived from our meta‐analysis a key group of organisms that can be used to identify individuals at risk of developing dental caries based on oral microbiome inhabitants.

OBJECTIVES:There are several hypotheses regarding how chlorhexidine (CHX) digluconate causes staining with the role of beverages, specifically the precipitation of anionic dietary chromogens onto adsorbed cations, the most probable cause. The aim of this study was to investigate and compare the staining potential of common beverages using an in vitro staining and brushing model to better understand the interactions between chromogens from different beverage categories and the teeth.MATERIALS AND METHODS:Human enamel samples were exposed to a cyclic treatment of artificial saliva and 0.2% CHX mouthwash combined with a range of beverages, with and without brushing, simulating a period equivalent to 2 weeks. Eleven beverages were tested: diet coke, diet lemonade, white wine, red wine, lager beer, black tea, coffee, black tea with milk, coffee with milk, ginger and lemon infusion, and water. Toothbrushing was performed in a brushing simulator with toothpaste and also with water. Colorimetric differences were determined by ΔE using a VITA Easyshade dental spectrophotometer. Statistical analyses were performed by one-way analysis of variance with post hoc Tukey's honestly significant difference test and Levene's test.RESULTS:Black tea and red wine produced highest staining, which agrees with the literature. Significant staining was also observed for a ginger and lemon infusion, coffee, coffee with milk, tea with milk, and lager beer compared with water (p < 0.05). The staining potential of diet coke in combination with brushing appeared to be connected to its low pH. Both white wine and diet lemonade produced stain comparable to the water control. After treatment with high staining beverages, scanning electron microscope evaluation confirmed the formation of a surface layer. The mechanical resistance of the stain differed depending on the beverage, black tea stain was the most resistant. The addition of milk to tea and coffee considerably modified the stain layer and the adhesion to the tooth surface.CONCLUSION:The data may help demonstrate that appropriate user guidance can avoid stain and in turn help improve user compliance during short-term use of this gold standard antimicrobial treatment.

Denture stomatitis (DS) is an inflammatory disease resulting from a polymicrobial biofilm perturbation at the denture surface–palatal mucosa interface. Recommendations made by dental health care professionals often lack clarity for appropriate denture cleaning. This study investigated the efficacy of brushing with off-the-shelf denture cleanser (DC) tablets (Poligrip®) vs. two toothpastes (Colgate® and Crest®) in alleviating the viable microorganisms (bacteria and fungi) in an in vitro denture biofilm model. Biofilms were grown on poly(methyl)methacrylate (PMMA) discs, then treated daily for 7 days with mechanical disruption (brushing), plus Poligrip® DC, Colgate® or Crest® toothpastes. Weekly treatment with Poligrip® DC on day 7 only was compared to daily modalities. All treatment parameters were processed to determine viable colony forming units for bacteria and fungi using the Miles and Misra technique, and imaged by confocal laser scanning microscopy (CLSM). Brushing with daily DC therapy was the most effective treatment in reducing the viable biofilm over 7 days of treatment. Brushing only was ineffective in controlling the viable bioburden, which was confirmed by CLSM imaging. This data indicates that regular cleansing of PMMA with DC was best for polymicrobial biofilms.

Background: Existing standardized biofilm assays focus on simple mono-species or bacterial-only models. Incorporating Candida albicans into complex biofilm models can offer a more appropriate and relevant polymicrobial biofilm for the development of oral health products. Aims: This study aimed to assess the importance of interkingdom interactions in polymicrobial oral biofilm systems with or without C. albicans, and test how these models respond to oral therapeutic challenges in vitro. Materials and Methods: Polymicrobial biofilms (two models containing 5 and 10 bacterial species, respectively) were created in parallel in the presence and absence of C. albicans and challenged using clinically relevant antimicrobials. The metabolic profiles and biomasses of these complex biofilms were estimated using resazurin dye and crystal violet stain, respectively. Quantitative PCR was utilized to assess compositional changes in microbial load. Additional assays, for measurements of pH and lactate, were included to monitor fluctuations in virulence “biomarkers.” Results: An increased level of metabolic activity and biomass in the presence of C. albicans was observed. Bacterial load was increased by more than a factor of 10 in the presence of C. albicans. Assays showed inclusion of C. albicans impacted the biofilm virulence profiles. C. albicans did not affect the biofilms’ responses to the short-term incubations with different treatments. Conclusions: The interkingdom biofilms described herein are structurally robust and exhibit all the hallmarks of a reproducible model. To our knowledge, these data are the first to test the hypothesis that yeasts may act as potential “keystone” components of oral biofilms.

Objectives The aims of this study were to (1) investigate the relative time-dependent disruption and bactericidal effects of detergent-type surfactants on single- or dual-species biofilms of root canal isolates and (2) to examine the utility of polygonal graphs for depiction of biofilm disruption and cell killing. Materials and methods Single-species biofilms of Streptococcus sanguinis , Enterococcus faecalis , Fusobacterium nucleatum and Porphyromonas gingivalis were grown on nitro-cellulose membranes for 72 h and immersed in Tween®80, cetyltrimethylammoniumbromide (CTAB), and sodium dodecyl sulphate (SDS) for 1-, 5- or 10-min ( n = 3 per test). The number of viable and non-viable bacteria “disrupted” from the biofilm and those “remaining-attached” was determined using a viability stain in conjunction with fluorescence microscopy. The data were analysed using non-parametric Kruskal-Wallis test with 5% significance level. Results Gram-negative obligate anaerobes were more susceptible to cell removal than gram-positive facultative anaerobes. The majority of cells were disrupted after 1-min of exposure; however, the extent varied according to the agent and species. CTAB and SDS were more effective than Tween 80™ at disrupting biofilms and killing cells but all agents failed to achieve 100% disruption/kill. Conclusions Biofilm disruption and cell viability were influenced by the species, the test agent and the duration of exposure. CTAB and SDS were more effective in biofilm disruption than Tween 80™. Graphical depiction of biofilm disruption- and viability-outcomes provides an alternative means of simultaneously visualising and analysing relative efficacy in different domains. Clinical relevance Surfactants were not as effective at biofilm disruption as NaOCl but may be added to other non-disruptive antibacterial agents to enhance this property.

Photodynamic therapy (PDT) has been proposed as a new technique to inactivate microorganisms as it does not lead to the selection of mutant resistant strains; a clear benefit compared to antibiotic treatment. PDT has also attracted the interest of nanotechnology as the effectiveness of the treatment can be greatly enhanced by the use of nanoparticles. In the last decade, different approaches to the combination of nanoparticles and PDT have been investigated in relation to the antimicrobial applications of the technique. One use of the nanoparticles is to improve the delivery of photosensitiser to the bacteria; others use the nanoparticles to improve the inactivation kinetics. A different approach utilises nanoparticles as a photosensitiser. In this review these diverse types of interactions will be described.

Key to onset and progression of periodontitis is a complex relationship between oral bacteria and the host. The organisms most associated with severe periodontitis are the periodontal pathogens of the red complex: Tannerella forsythia, Treponema denticola and Porphyromonas gingivalis. These organisms express sialidases, which cleave sialic acid from host glycoproteins, and contribute to disease through various mechanisms. Here, we expressed and purified recombinant P. gingivalis sialidase SiaPG (PG_0352) and characterized its activity on a number of substrates, including host sialoglycoproteins and highlighting the inability to cleave diacetylated sialic acids - a phenomenon overcome by the NanS sialate-esterase from T. forsythia. Indeed SiaPG required NanS to maximize sialic acid harvesting from heavily O-acetylated substrates such as bovine salivary mucin, hinting at the possibility of interspecies cooperation in sialic acid release from host sources by these members of the oral microbiota. Activity of SiaPG and P. gingivalis was inhibited using the commercially available chemotherapeutic zanamivir, indicating its potential as a virulence inhibitor, which also inhibited sialic acid release from mucin, and was capable of inhibiting biofilm formation of P. gingivalis on oral glycoprotein sources. Zanamivir also inhibited attachment and invasion of oral epithelial cells by P. gingivalis and other periodontal pathogens, both in monospecies but also in multispecies infection experiments, indicating potential to suppress host-pathogen interactions of a mixed microbial community. This study broadens our understanding of the multifarious roles of bacterial sialidases in virulence, and indicates that their inhibition with chemotherapeutics could be a promising strategy for periodontitis therapy.

OBJECTIVES:Short-term use of sodium bicarbonate (NaHCO3)-containing toothpaste reduces plaque and improves clinical measures of gingivitis. To examine this over a longer period, we compared efficacy and tolerability of twice-daily brushing for 24 weeks with 67% or 0% NaHCO3-containing toothpastes in USA-based participants with moderate gingivitis (Clinicaltrials.gov:NCT02207400).METHODS:This was a six-month, randomized, examiner-blind, parallel-group, clinical trial. Investigators randomized adults with blood in expectorate after brushing and ≥ 20 gingival bleeding sites to 67% NaHCO3 (n = 123; n = 107 completed study) or 0% NaHCO3 (n = 123; n = 109 completed study) toothpastes. Primary efficacy variables included between-treatment differences in number of bleeding sites and Modified Gingival Index (MGI) score at 24 weeks. Secondary efficacy variables included Bleeding Index and Turesky modification of the Quigley-Hein Plaque Index (overall and interproximal sites) at six, 12, and 24 weeks. A subset of 50 participants underwent sampling to assess plaque microbiology over the course of treatment.RESULTS:Compared with the 0% NaHCO3 toothpaste, the 67% NaHCO3 toothpaste produced statistically significant improvements at Week 24 in number of bleeding sites (46.7% difference) and MGI (33.9% difference), and for all other endpoints (all p < 0.0001). There was no significant between-treatment difference in the proportion of participants harboring opportunistic pathogens. Products were generally well tolerated, with two and five treatment-related adverse events reported in the 67% and 0% NaHCO3 toothpaste groups, respectively.CONCLUSIONS:Gingival bleeding, gingivitis, and plaque indices were significantly improved at six, 12, and 24 weeks with twice-daily brushing with 67% NaHCO3-containing toothpaste in participants with moderate gingivitis.

The present study investigated the compounds present in the low molecular mass fraction of Lentinus edodes mushroom (shiitake) extract and their anti-virulence activity against oral pathogens (reference and clinical Streptococcus mutans, Actinomyces naeslundii, and Prevotella intermedia strains). Oxalic, succinic, and quinic acids, and adenine, inosine, and uridine were identified by HPLC-DAD-ESI-MS/MS. Their anti-biofilm production and preformed biofilm disaggregation activities were studied using commercial standard compounds at different concentrations. As regards S. mutans, the highest activity was shown by adenine at 5 mg mL-1 both in the biofilm inhibition (BI 50%) and biofilm disaggregation tests (BD 20%). Considering A. naeslundii, BI values close to 80% were registered for oxalic acid at 1 mg mL-1 and 2 mg mL-1 and BD 50% for quinic acid at 3 mg mL-1. A weaker activity was found against P. intermedia. Furthermore, different mixtures of the commercial standards were tested showing that the activity of a compound can be strongly and sometimes negatively affected by the presence of the other compounds.

Bacterial sialidases cleave terminal sialic acid from a variety of host glycoproteins, and contribute to survival and growth of many human-dwelling bacterial species, including various pathogens. Tannerella forsythia, an oral, Gram-negative, fastidious anaerobe, is a key organism in periodontal disease and possesses a dedicated sialic acid utilisation and scavenging (nan) operon, including NanH sialidase. Here, we describe biochemical characterisation of recombinant NanH, including its action on host-relevant sialoglycans such as sialyl Lewis A and sialyl Lewis X (SLeA/X), and on human cell-attached sialic acids directly, uncovering that it is a highly active broad specificity sialidase. Furthermore, the N-terminal domain of NanH was hypothesised and proved to be capable of binding to a range of sialoglycans and non-sialylated derivatives with Kd in the micromolar range, as determined by steady-state tryptophan fluorescence spectroscopy, but it has no catalytic activity in isolation from the active site. We consider this domain to represent the founding member of a novel subfamily of carbohydrate-binding module (CBM), involved in glycosidase-ligand binding. In addition, we created a catalytically inactive version of the NanH enzyme (FRIP → YMAP) that retained its ability to bind sialic acid-containing ligands and revealed for the first time that binding activity of a CBM is enhanced by association with the catalytic domain. Finally, we investigated the importance of Lewis-type sialoglycans on T. forsythia-host interactions, showing that nanomolar amounts of SLeA/X were capable of reducing invasion of oral epithelial cells by T. forsythia, suggesting that these are key ligands for bacterial-cellular interactions during periodontal disease.

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Denture-associated stomatitis is a common candidal infection that may give rise to painful oral symptoms, as well as be a reservoir for infection at other sites of the body. As poly (methyl methacrylate) (PMMA) remains the main material employed in the fabrication of dentures, the aim of this research was to evaluate the adhesion of Candida albicans cells onto PMMA surfaces by employing an atomic force microscopy (AFM) single-cell force spectroscopy (SCFS) technique. For experiments, tipless AFM cantilevers were functionalized with PMMA microspheres and probed against C. albicans cells immobilized onto biopolymer-coated substrates. Both a laboratory strain and a clinical isolate of C. albicans were used for SCFS experiments. Scanning electron microscopy (SEM) and AFM imaging of C. albicans confirmed the polymorphic behavior of both strains, which was dependent on growth culture conditions. AFM force-spectroscopy results showed that the adhesion of C. albicans to PMMA is morphology dependent, as hyphal tubes had increased adhesion compared with yeast cells (P < 0.05). C. albicans budding mother cells were found to be nonadherent, which contrasts with the increased adhesion observed in the tube region. Comparison between strains demonstrated increased adhesion forces for a clinical isolate compared with the lab strain. The clinical isolate also had increased survival in blood and reduced sensitivity to complement opsonization, providing additional evidence of strain-dependent differences in Candida-host interactions that may affect virulence. In conclusion, PMMA-modified AFM probes have shown to be a reliable technique to characterize the adhesion of C. albicans to acrylic surfaces.

This in vitro study evaluated the effects of polishing the fitting surface of two acrylic based matetrials on the surface characteristics and biofilm formation of Methicillin Resistance Staphylococcus aureus (MRSA) biofilms. A simulated mouth model (Constant Depth Film Fermenter) was used to produce biofilms on autopolymerised and heat cure acrylic substrata. Surface parameters included surface roughness, hydrophobicity and surface free energy was evaluated after using the conventional polishing procedure to samples simulating the clinical situation. The results showed that MRSA has successfully adhered and grown in all samples with a similar pattern without a significant difference between the recovered viable bacterial counts. The rough surface provide protect- ed niche for MRSA against the dislodgment forces. In conclusion, the finding of suggested that polishing the fitting surface of acrylic base materials may facilitate MRSA biofilm removal.

ObjectivesDental implants are prone to failure as a result of bacterial biofilm accumulation. Such biofilms are often resistant to traditional antimicrobials and the use of nanoparticles as implant coatings may offer a means to control infection over a prolonged period. The objective of this study was to determine the antibiofilm activity of nanoparticulate coated titanium (Ti) discs using a film fermenter based system.MethodsMetal oxide nanoparticles of zinc oxide (nZnO), hydroxyapatite (nHA) and a combination (nZnO+nHA) were coated using electrohydrodynamic deposition onto Ti discs. Using human saliva as an inoculum, biofilms were grown on coated discs for 96h in a constant depth film fermenter under aerobic conditions with artificial saliva and peri-implant sulcular fluid. Viability assays and biofilm thickness measurements were used to assess antimicrobial activity.ResultsFollowing 96h, reduced numbers of facultatively anaerobic and Streptococcus spp. on all three nano-coated surfaces were demonstrated. The proportion of non-viable microorganisms was shown to be higher on nZnO and composite (nZnO+nHA) coated surfaces at 96h compared with nHA coated and uncoated titanium. Biofilm thickness comparison also demonstrated that nZnO and composite coatings to be the most effective.ConclusionsThe findings support the use of coating Ti dental implant surfaces with nZnO to provide an antimicrobial function.Clinical significanceCurrent forms of treatment for implant associated infection are often inadequate and may result in chronic infection requiring implant removal and resective/regenerative procedures to restore and reshape supporting tissue. The use of metal oxide nanoparticles to coat implants could provide osteoconductive and antimicrobial functionalities to prevent failure.

Biofilm development on the internal surfaces of dental implants and dental implant components may be associated with peri-implant hard and soft tissue abnormalities. Currently, no protocols have been established for effective disinfection of removable implant components. The purpose of this study was to evaluate the antibiofilm properties of commonly used dental antimicrobial agents (chlorhexidine and delmopinol) on multispecies biofilms. Biofilms of 48 h and 168 h maturity were exposed to 0.2% chlorhexidine, 2% chlorhexidine and a product containing 0.2% delmopinol for 5, 10 and 20 min. 2% chlorhexidine was the most effective agent, achieving a total viable biofilm reduction ranging from 96.2% to >99.99% depending on the time of exposure and the stage of biofilm development.

OBJECTIVES:Sodium bicarbonate has been shown clinically to be efficacious at removing dental plaque; however, its effect of mechanism against biofilms has not been evaluated in vitro. Here, we used a well-established in vitro plaque biofilm model to investigate the disruption of dental plaque biofilms.METHODS:Biofilms were grown in a constant depth film fermentor for up to 14 days. The fermentor was inoculated with pooled human saliva and growth maintained with artificial saliva. After various time points, replicate biofilms were removed and subjected to treatment at varying concentrations of sodium bicarbonate. Disruption of the plaque was assessed by viable counts and microscopy.RESULTS:The viable count results showed that younger biofilms were less susceptible to the action of sodium bicarbonate; however, biofilms of 7 days and older were increasingly susceptible to the material with the oldest biofilms being the most susceptible. Sixty-seven percentage of sodium bicarbonate slurry was able to reduce the number of organisms present by approx. 3 log10 . These quantitative data were corroborated qualitatively with both confocal and electron microscopy, which both showed substantial qualitative removal of mature biofilms.CONCLUSIONS:The results from this study have shown that sodium bicarbonate is able to disrupt mature dental plaque grown in vitro and that its reported efficacy in maintaining oral hygiene may be related to this key factor.

Staphylococci species have been isolated from removable orthodontic retainers. The aims of this study were to determine the most suitable device to analyze surface roughness of autopolymerized acrylic and thermoplastic materials and whether the surface dynamics of these materials influences the attachment of Methicillin-Resistant Staphylococcus aureus (MRSA). Clinically simulated samples of autopolymerized acrylic and thermoplastic material were first evaluated using laser non-contact, stylus mechanical profilometries and atomic force microscopy (AFM) followed by contact angle measurement to characterize their surface dynamics. Finally, an in vitro biofilm assay was carried out using a constant depth film fermentor to assess biofilm attachment. The results showed a significant difference between the roughness values obtained from the tested profilometers with the AFM exhibiting the most consistent roughness values. MRSA tended to accumulate initially within the microscopic irregularities of autopolymerized acrylic samples whereas acid-base and electron donor interactions influenced the bacterial attachment onto the thermoplastic samples.