Assessment of Faecal Microbiota Transplant Stability in Deep-Freeze Conditions: A 12-Month Ex Vivo Viability Analysis.

BACKGROUND: Faecal microbiota transplantation (FMT) is an established treatment for Clostridioides difficile infection and is under investigation for other conditions. The availability of suitable donors and the logistics of fresh stool preparation present challenges, making frozen, biobanked stools an attractive alternative. AIMS: This study aimed to evaluate the long-term viability of bacterial populations in faecal samples stored at -80°C for up to 12 months, supporting the feasibility of using frozen grafts for FMT. METHODS: Fifteen faecal samples from nine healthy donors were processed, mixed with cryoprotectants and stored at -80°C. Samples were assessed at baseline and after 3, 6 and 12 months using quantitative culturing methods to determine the concentration of live bacteria. RESULTS: Quantitative analysis showed no significant decrease in bacterial viability over the 12-month period for both aerobic and anaerobic cultures (p = 0.09). At all timepoints, the coefficients of variability in colony-forming unit (CFU) counts were greater between samples (102 ± 21% and 100 ± 13% for aerobic and anaerobic cultures, respectively) than the variability between measurements of the same sample (30 ± 22% and 30 ± 19%). CONCLUSIONS: The study confirmed that faecal microbiota can be preserved with high viability in deep-freeze storage for up to a year, making allogenic FMT from biobanked samples a viable and safer option for patients. However, a multidonor approach may be beneficial to mitigate the risk of viability loss in any single donor sample.

Development of a novel in vitro model for non-cavitated carious lesion formation reflecting carious lesion activity.

BACKGROUND: Oral biofilms are a critical component in dental caries formation. However, current remineralization studies often overlook the impact of microbial factors. Therefore, a comprehensive clinically relevant assessment of caries is needed. This study aimed to develop a novel in vitro model capable of generating non-cavitated carious lesions that incorporates both mineral loss and microbial activity using quantitative light-induced fluorescence-digital (QLF-D) technology. METHODS: A total of 44 artificial early carious lesions were formed using bovine incisors. The extent of fluorescence loss (ΔF) was analyzed using a QLF-D camera. Oral microcosm biofilms were then employed to construct 22 active and 22 inactive carious lesions. The red fluorescence emission rate (ΔR) and bacterial viability (RatioG/G+R) was measured using QLF-D camera and a live-dead bacterial assay, respectively. Independent t-tests were performed to compare ΔF, ΔR, and bacterial viability of artificial carious lesions according to their activity status. RESULTS: No significant difference in ΔF between the lesions was found based on activity status (p = 0.361). However, the ΔR of active lesions was 1.82 times higher than that of inactive lesions, and the RatioG/G+R was 1.49 times higher in active lesions than in inactive lesions (both p < 0.001). CONCLUSIONS: The significant differences observed in ΔR and RatioG/G+R between active and inactive lesions emphasize the importance of considering lesion activity status when evaluating the potential efficacy of remineralization agents. This study presents a novel in vitro remineralization assessment model that reflects carious lesion activity while controlling baseline mineral distributions of lesions.

Rapid detection of viable microbes with 5-cyano-2,3-di-(p-tolyl)tetrazolium chloride and 5(6)-carboxyfluorescein diacetate using a fibre fluorescence spectroscopy system.

AIMS: To assess the efficacy of two commercially available viability dyes, 5-cyano-2,3-di-(p-tolyl)tetrazolium chloride (CTC) and 5(6)-carboxyfluorescein diacetate (CFDA), in reporting on viable cell concentration and species using an all-fibre fluorometer. METHODS AND RESULTS: Four bacterial species (two Gram-positive and two Gram-negative) commonly associated with food poisoning or food spoilage (Escherichia coli, Salmonella enterica, Staphylococcus aureus, and Bacillus cereus) were stained with CTC or CFDA and the fibre fluorometer was used to collect full fluorescence emission spectra. A good correlation between concentration and fluorescence intensity was found for Gram-negative bacteria between 107 and 108 colony-forming units (CFU) ml-1. There was no correlation with concentration for Gram-positive bacteria; however, the information in the CTC and CFDA spectra shows the potential to distinguish Gram-negative cells from Gram-positive cells, although it may simply reflect the overall bacterial metabolic activity under staining conditions from this study. CONCLUSIONS: The limit of detection (LoD) is too high in the dip-probe approach for analysis; however, the development of an approach measuring the fluorescence of single cells may improve this limitation. The development of new bacteria-specific fluorogenic dyes may also address this limitation. The ability to differentiate bacteria using these dyes may add value to measurements made to enumerate bacteria using CTC and CFDA.

Effectiveness of Umonium38 against Burkholderia pseudomallei, Escherichia coli, Pseudomonas aeruginosa and Methicillin-Resistant Staphylococcus aureus (MRSA).

AIMS: We investigated the antibacterial efficacy of Umonium38 and Virkon® against Burkholderia pseudomallei, Escherichia coli, Pseudomonas aeruginosa and Methicillin-Resistant Staphylococcus aureus (MRSA) up to 14 days following treatment. METHODS AND RESULTS: Umonium38 was diluted to 0.5%, 1.0%, 1.5%, 2.0%, 2.5% and 3%, tested against the bacterial strains at various contact times (15 min to 24 h), and incubated for up to 14 days. A minimum concentration of 0.5% Umonium38 with a contact time of 15 min effectively killed approximately 108 CFU/ml of all four bacterial species. No growth was observed on agar plates from day 0 until day 14 for all six concentrations. The bacteria were also inactivated by a 30-minute treatment time using Virkon® 1% solution. CONCLUSIONS: Umonium38 effectively inactivates B. pseudomallei, E. coli, P. aeruginosa and MRSA at a concentration of ≥ 0.5% with a contact time of at least 15 min. The antimicrobial effect of Umonium38 remained for 14 days.

Ferrous sulfide nanoparticles can be biosynthesized by sulfate-reducing bacteria: Synthesis, characterization and removal of heavy metals from acid mine drainage.

Ferrous sulfide nanoparticles (nFeS) have proven to be effective in removing heavy metals (HMs) from wastewater. One such approach, which has garnered much attention as a sustainable technology, is via the in situ microbial synthesis of nFeS. Here, a sulfate-reducing bacteria (SRB) strain, Geobacter sulfurreducens, was used to initially biosynthesize ferrous sulfide nanoparticles (SRB-nFeS) and thereafter remove HMs from acid mine drainage (AMD). SRB-nFeS was characterized by X-ray powder diffraction (XRD), scanning electron microscopy (SEM) coupled to an energy dispersive spectrometer (EDS), three-dimensional excitation-emission matrix (3D-EEM) spectroscopy, Fourier transform infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS). Such characterization showed that SRB mediated the reduction of SO42- to S2- to form nFeS, where the metabolized substances functioned as complexing agents which coordinated with nFeS to form biofunctional SRB-nFeS with improved stability. One advantage of this synthetic route was that the attachment of nFeS to the bacterial surface protected SRB cells from HM toxicity. Furthermore, due to a synergistic effect between nFeS and SRB, HM removal from both solution and AMD by SRB-nFeS was enhanced relative to the constituent components. Thus, after 5 consecutive cycles of HM removal, SRB-nFeS removed, Pb(Ⅱ) (92.6%), Cd(Ⅱ) (78.7%), Cu(Ⅱ) (76.0%), Ni(Ⅱ) (62.5%), Mn(Ⅱ) (62.2%), and Zn(Ⅱ) (88.5%) from AMD This study thus provides new insights into the biosynthesis of SRB-nFeS and its subsequent practical application in the removal of HMs from AMD.

Bacterial viability in dry-surface biofilms in healthcare facilities: a systematic review.

BACKGROUND: Bacteria are known to live inside architectural structures called biofilms. Though standard biofilms have been studied extensively for more than 50 years, little is known about dry-surface biofilms (DSBs). Since 2012, DSBs have been described in several scientific papers, but basic knowledge about the viability and culturability of bacteria remains limited. AIM: To conduct a systematic review to determine whether bacteria inside DSBs are viable, culturable, and enumerable. METHODS: Eligible articles had to deal with DSBs containing at least one bacterial species involved in healthcare-associated infections, which developed in actual healthcare environments (in-situ) or with the help of any biofilm model (in-vitro). FINDINGS: Twenty-four articles were included in the review. Whereas most of them isolated viable bacteria (87% in situ; 100% in vitro), no in-situ study quantified culturable bacteria in the biofilm per unit area. Conversely, 100% of in-vitro studies cultured the bacteria from controls and 94.4% supplied an enumeration of them. Culturable bacteria also grew after 78% of the cleaning, disinfection, or sterilization protocols tested. Microscopic observations after staining the samples with live/dead fluorescent probes (Baclight®) showed large amounts of viable cells in culture-negative samples. CONCLUSION: Our study questions the efficacy of current methods for microbiological monitoring of surfaces, since these methods are only based on bacterial culturability. To improve both surface monitoring and cleaning and disinfection protocols, it is necessary to integrate the concept of DSBs which appears to contain a significant amount of viable but non-culturable bacteria.

Antibacterial Activity of Root Repair Cements in Contact with Dentin-An Ex Vivo Study.

This study assessed the antibacterial characteristics of the dentin/material interface and dentin surfaces exposed to experimental hydraulic calcium silicate cement (HCSC) with or without bioactive glass (BG) replacement (20% or 40%) or mixed with a silver nanoparticle (SNP) solution (1 or 2 mg/mL), and Biodentine, TotalFill BC RRM putty and Intermediate Restorative Material (IRM). Human root dentin segments with test materials were assessed at 1 or 28 days. In one series, the specimens were split to expose the dentin and material surfaces. A 24 h direct contact test was conducted against three-day established Enterococcus faecalis and Pseudomonas aeruginosa monospecies biofilms. In another series, the dentin/material interface of intact specimens was exposed to biofilm membranes for 3 days and the antibacterial activity was assessed via confocal microscopy. The interface was additionally characterised. All one-day material and dentin surfaces were antibacterial. Dentin surfaces exposed to HCSC with 40% BG-replacement, Biodentine and IRM had decreased antibacterial properties compared to those of the other cements. The HCSC mixed with a 2 mg/mL SNP solution had the highest antimicrobial effect in the confocal assay. The interfacial characteristics of HCSCs were similar. The test materials conferred antibacterial activity onto the adjacent dentin. The BG reduced the antibacterial effect of dentin exposed to HCSC; a 2 mg/mL SNP solution increased the antibacterial potential for longer interaction periods (three-day exposure).

Anaerobic Feces Processing for Fecal Microbiota Transplantation Improves Viability of Obligate Anaerobes.

Fecal microbiota transplantation (FMT) is under investigation for several indications, including ulcerative colitis (UC). The clinical success of FMT depends partly on the engraftment of viable bacteria. Because the vast majority of human gut microbiota consists of anaerobes, the currently used aerobic processing protocols of donor stool may diminish the bacterial viability of transplanted material. This study assessed the effect of four processing techniques for donor stool (i.e., anaerobic and aerobic, both direct processing and after temporary cool storage) on bacterial viability. By combining anaerobic culturing on customized media for anaerobes with 16S rRNA sequencing, we could successfully culture and identify the majority of the bacteria present in raw fecal suspensions. We show that direct anaerobic processing of donor stool is superior to aerobic processing conditions for preserving the bacterial viability of obligate anaerobes and butyrate-producing bacteria related to the clinical response to FMT in ulcerative colitis patients, including Faecalibacterium, Eubacterium hallii, and Blautia. The effect of oxygen exposure during stool processing decreased when the samples were stored long-term. Our results confirm the importance of sample conditioning to preserve the bacterial viability of oxygen-sensitive gut bacteria. Anaerobic processing of donor stool may lead to increased clinical success of FMT, which should further be investigated in clinical trials.

Qualitative and Quantitative Methods to Measure Antibacterial Activity Resulting from Bacterial Competition.

In the environment, bacteria compete for niche occupancy and resources; they have, therefore, evolved a broad variety of antibacterial weapons to destroy competitors. Current laboratory techniques to evaluate antibacterial activity are usually labor intensive, low throughput, costly, and time consuming. Typical assays rely on the outgrowth of colonies of prey cells on selective solid media after competition. Here, we present fast, inexpensive, and complementary optimized protocols to qualitatively and quantitively measure antibacterial activity. The first method is based on the degradation of a cell-impermeable chromogenic substrate of the ß-galactosidase, a cytoplasmic enzyme released during lysis of the attacked reporter strain. The second method relies on the lag time required for the attacked cells to reach a defined optical density after the competition, which is directly dependent on the initial number of surviving cells. Key features First method utilizes the release of ß-galactosidase as a proxy for bacterial lysis. Second method is based on the growth timing of surviving cells. Combination of two methods discriminates between cell death and lysis, cell death without lysis, or survival to quasi-lysis. Methods optimized to various bacterial species such as Escherichia coli, Pseudomonas aeruginosa, and Myxococcus xanthus. Graphical overview.

Bacterial DNA on the skin surface overrepresents the viable skin microbiome.

The skin microbiome provides vital contributions to human health. However, the spatial organization and viability of its bacterial components remain unclear. Here, we apply culturing, imaging, and molecular approaches to human and mouse skin samples, and find that the skin surface is colonized by fewer viable bacteria than predicted by bacterial DNA levels. Instead, viable skin-associated bacteria are predominantly located in hair follicles and other cutaneous invaginations. Furthermore, we show that the skin microbiome has a uniquely low fraction of viable bacteria compared to other human microbiome sites, indicating that most bacterial DNA on the skin surface is not associated with viable cells Additionally, a small number of bacterial families dominate each skin site and traditional sequencing methods overestimate both the richness and diversity of the skin microbiome. Finally, we performed an in vivo skin microbiome perturbation-recovery study using human volunteers. Bacterial 16S rRNA gene sequencing revealed that, while the skin microbiome is remarkably stable even in the wake of aggressive perturbation, repopulation of the skin surface is driven by the underlying viable population. Our findings help explain the dynamics of skin microbiome perturbation as bacterial DNA on the skin surface can be transiently perturbed but is replenished by a stable underlying viable population. These results address multiple outstanding questions in skin microbiome biology with significant implications for future efforts to study and manipulate it.

Classification between live and dead foodborne bacteria with hyperspectral microscope imagery and machine learning.

Identifying live foodborne bacteria is essential for ensuring food safety and preventing foodborne illnesses. This study investigated the use of hyperspectral microscope imaging and deep learning methods to accurately distinguish between live and dead foodborne bacteria based on their spectral and morphological features. Three deep learning models, Fusion-Net I, II, and III, were developed and evaluated for their ability to classify live and dead bacterial cells of six pathogenic strains, including Escherichia coli (EC), Listeria innocua (LI), Staphylococcus aureus (SA), Salmonella Enteritidis (SE), Salmonella Heidelberg (SH), and Salmonella Typhimurium (ST). The models utilized both morphological and spectral characteristics of the bacterial cells, with inputs of average spectra and 546 nm band images. Fusion-Net I achieved high accuracy in identifying live bacterial cells, with a classification accuracy of 100% for LI, SE, ST strains and over 92.9% for EC, SA, SH. Fusion-Net II and III models were even more robust, achieving 100% accuracy consistently in classifying dead cells in all six strains. Fusion-Net III also showed the ability to identify bacterial strains with 96.9% accuracy, making it a dual-task model with potential applications in identifying live foodborne bacteria prior to foodborne outbreaks. These findings suggest that the use of hyperspectral microscope imaging and deep learning methods could provide a new tool for quickly and accurately identifying bacterial viability, thereby improving the efficiency and reliability of food safety inspection.

Dry alginate beads for fecal microbiota transplantation: From model strains to fecal samples.

Clostridioides difficile infection (CDI) is a critical nosocomial infection with more than 124,000 cases per year in Europe and a mortality rate of 15-17 %. The standard of care (SoC) is antibiotic treatment. Unfortunately, the relapse rate is high (∼35 %) and SoC is significantly less effective against recurrent infection (rCDI). Fecal microbiota transplantation (FMT) is a recommended treatment against rCDI from the second recurrence episode and has an efficacy of 90 %. The formulation of diluted donor stool deserves innovation because its actual administration routes deserve optimization (naso-duodenal/jejunal tubes, colonoscopy, enema or several voluminous oral capsules). Encapsulation of model bacteria strains in gel beads were first investigated. Then, the encapsulation method was applied to diluted stools. Robust spherical gel beads were obtained. The mean particle size was around 2 mm. A high loading of viable microorganisms was obtained for model strains and fecal samples. For plate-counting, values ranged from 1015 to 1017 CFU/g for single and mixed model strains, and 106 to 108 CFU/g for fecal samples. This corresponded to a viability of 30 % to 60 % as assessed by flow cytometry. This novel formulation is promising as the technology is applicable to both model strains and bacteria contained in the gut microbiota.

The Survival of Psychobiotics in Fermented Food and the Gastrointestinal Tract: A Review.

In recent years, scientists have been particularly interested in the gut-brain axis, as well as the impact of probiotics on the nervous system. This has led to the creation of the concept of psychobiotics. The present review describes the mechanisms of action of psychobiotics, their use in food products, and their viability and survival during gastrointestinal passage. Fermented foods have a high potential of delivering probiotic strains, including psychobiotic ones. However, it is important that the micro-organisms remain viable in concentrations ranging from about 106 to 109 CFU/mL during processing, storage, and digestion. Reports indicate that a wide variety of dairy and plant-based products can be effective carriers for psychobiotics. Nonetheless, bacterial viability is closely related to the type of food matrix and the micro-organism strain. Studies conducted in laboratory conditions have shown promising results in terms of the therapeutic properties and viability of probiotics. Because human research in this field is still limited, it is necessary to broaden our understanding of the survival of probiotic strains in the human digestive tract, their resistance to gastric and pancreatic enzymes, and their ability to colonize the microbiota.

A PMAxxTM qPCR Assay Reveals That Dietary Administration of the Microalgae Tetraselmis chuii Does Not Affect Salmonella Infantis Caecal Content in Early-Treated Broiler Chickens.

Salmonella enterica serovars cause infections in humans. S. enterica subsp. enterica serovar Infantis is considered relevant and is commonly reported in poultry products. Evaluating innovative approaches for resisting colonization in animals could contribute to the goal of reducing potential human infections. Microalgae represent a source of molecules associated with performance and health improvement in chickens. Tetraselmis chuii synthesizes fermentable polysaccharides as part of their cell wall content; these sugars are known for influencing caecal bacterial diversity. We hypothesized if its dietary administration could exert a positive effect on caecal microbiota in favor of a reduced S. Infantis load. A total of 72 one-day-old broiler chickens (COBB 500) were randomly allocated into three groups: a control, a group infected with bacteria (day 4), and a group challenged with S. Infantis but fed a microalgae-based diet. Caecal samples (n = 8) were collected two days post-infection. A PMAxxTM-based qPCR approach was developed to assess differences regarding bacterial viable load between groups. The inclusion of the microalga did not modify S. Infantis content, although the assay proved to be efficient, sensitive, and repeatable. The utilized scheme could serve as a foundation for developing novel PCR-based methodologies for estimating Salmonella colonization.

Cryopreservation of stool samples altered the microbial viability quantitively and compositionally.

Stool is the most commonly used sample for gut microbiota analysis in humans and animals. Cryopreservation of stool at - 80 °C is a feasible and simple method in clinics and researches, especially in large-scale cohort studies. However, the viability of bacteria in stool after freezing has yet well-demonstrated quantitatively and compositionally. This study determined the viable microbiota of samples under cryopreservation at - 80 °C, relative to fresh samples and that stored at ambient. Stool samples were collected from three healthy adults. Propidium monoazide treatment combined with quantitative PCR and 16S rRNA gene sequencing was performed to target viable microbiota. After freezing, the number of viable bacteria decreased, though inter-individual difference existed. Notably, the alpha diversity of viable microbiota after freezing did not change significantly, while its composition changed. Freezing significantly reduced the viable bacteria in Gram-negative genera of Bacteroidetes and Firmicutes, and proportionally increased Gram-positive bacteria in genera of Actinobacteria and Firmicutes, including Bifidobacterium, Collinsella and Blautia, implying that the cell envelope structure associated with the bacterial sensitivity to freezing. On the contrary, the room temperature storage not only decreased the number of viable bacteria, but also decreased the microbial alpha diversity, and remarkably enriched facultative anaerobes of Escherichia-Shigella, Enterococcus and Lactococcus, some of which are opportunistic pathogens. Our findings suggested that changes in viable microbiota in stool samples caused by cryopreservation should be paid enough attention for downstream utilization.

A preliminary investigation into bacterial viability using scanning electron microscopy-energy-dispersive X-ray analysis: The case of antibiotics.

The metabolic stages of bacterial development and viability under different stress conditions induced by disinfection, chemical treatments, temperature, or atmospheric changes have been thoroughly investigated. Here, we aim to evaluate early metabolic modifications in bacteria following induced stress, resulting in alterations to bacterial metabolism. A protocol was optimized for bacterial preparation using energy-dispersive X-ray (EDX) microanalysis coupled with scanning electron microscopy (SEM), followed by optimizing EDX data acquisition and analysis. We investigated different preparation methods aiming to detect modifications in the bacterial chemical composition at different states. We first investigated Escherichia coli, acquiring data from fresh bacteria, after heat shock, and after contact with 70% ethanol, in order to prove the feasibility of this new strategy. We then applied the new method to different bacterial species following 1 h of incubation with increasing doses of antibiotics used as a stress-inducing agent. Among the different materials tested aiming to avoiding interaction with bacterial metabolites, phosphorous-doped silicon wafers were selected for the slide preparation. The 15 kV acceleration voltage ensured all the chemical elements of interest were excited. A thick layer of bacterial culture was deposited on the silicon wafer providing information from multiple cells and intra-cellular composition. The EDX spectra of fresh, heat-killed, and alcohol-killed E. coli revealed important modifications in magnesium, potassium, and sodium. Those same alterations were detected when applying this strategy to bacteria exposed to antibiotics. Tests based on SEM-EDX acquisition systems would provide early predictions of the bacterial viability state in different conditions, yielding earlier results than culture.

Antibiotic Resistance Influences Growth Rates and Cross-Tolerance to Lactic Acid in Escherichia coli O157:H7 H1730.

Escherichia coli O157:H7-contaminated beef has been implicated in numerous foodborne outbreaks. Contamination occurs despite the use of antimicrobial interventions such as lactic acid (LA). In addition, resistance to antibiotics such as ampicillin and streptomycin among isolates has been frequently reported. The influence of antibiotic resistance (ABR) on growth rates and cross-tolerance of lettuce isolate E. coli O157:H7 H1730 to LA was evaluated. Antibiotic-resistant strain variants were generated by conferring resistance to either ampicillin (ampC) or streptomycin (strepC) or both ampicillin and streptomycin (ampC strepC) through incremental exposure to the antibiotics. Ampicillin resistance was also conferred by plasmid transformation to generate the ampP and ampP strepC strains. The minimum inhibitory concentration of LA on all the strains evaluated was 0.375% v/v. The lag phase duration of all strains except E. coli O157:H7 ampP strepC increased with increasing concentration of LA. The ampP strepC and ampC strains were most tolerant to 5% LA with declines in the cell population of 2.86 and 2.56 log CFU/mL, respectively (p < 0.05). The ampP strepC strain was the most tolerant when evaluated by the live/dead viability assay. The addition of the efflux pump inhibitor, carbonyl cyanide m-chlorophenylhydrazone, with 2.5% LA resulted in a significant increase in sensitivity in the no resistance (NR) wild-type and ampC strains, resulting in 6.62 and 6.65 log CFU/mL reduction, respectively, while the highly tolerant ampP strepC strain had a 2.90 log CFU/mL decrease. Tolerance to LA was significantly influenced by both the ABR profile of the strain and LA concentration. The results from this study indicate that E. coli O157:H7 strains with certain ABR profiles might be more tolerant to LA.

Selective Quantification of Erwinia amylovora Live Cells in Pome Fruit Tree Cankers by Viability Digital PCR.

The accurate assessment of Erwinia amylovora live cell populations in fire blight cankers by classic microbiology methods has major limitations. Some of them are the presence of competitive microbiota in samples that inhibit E. amylovora's growth and the release of toxic compounds by plant material during sample processing, which may hamper the pathogen's ability to form colonies on solid media. Digital PCR (dPCR) combined with the photo-reactive DNA-binding dye propidium monoazide (PMA) allows selective detection and quantification of live E. amylovora cells in woody samples while overcoming the constraints of culture-dependent methods. This work describes a reliable viability dPCR procedure to determine E. amylovora live cell concentrations in fire blight cankers from pome fruit trees. This protocol can be adapted for the analysis of other types of plant material and enables investigation of ecological, epidemiological, and management significance of cankers as a relatively underexplored part of the fire blight disease cycle.

A PMMA bone cement with improved antibacterial function and flexural strength.

A novel non-leaching antibacterial bone cement has been developed and evaluated. An antibacterial furanone derivative was synthesized and covalently coated onto the surface of alumina filler particles, followed by mixing into a conventional poly(methyl methacrylate) bone cement. Flexural strength and bacterial viability were used to evaluate the modified cements. Effects of coated antibacterial moiety content, coated alumina filler particle size and loading were investigated. Results showed that almost all the modified cements showed higher flexural strength (up to 10%), flexural modulus (up to 18%), and antibacterial activity (up to 67% to S. aureus and up to 84% to E. coli), as compared to original poly(methyl methacrylate) cement. Increasing antibacterial moiety and filler loading significantly enhanced antibacterial activity. On the other hand, increasing coated filler particle size decreased antibacterial activity. Increasing antibacterial moiety content and particle size did not significantly affect flexural strength and modulus. Increasing filler loading did not significantly affect flexural modulus but reduced flexural strength. Antibacterial agent leaching tests showed that it seems no leachable antibacterial component from the modified experimental cement to the surrounding environment. Within the limitations of this study, the modified poly(methyl methacrylate) bone cement may potentially be developed into a clinically useful bone cement for reducing in-surgical and post-surgical infection.

A novel antibacterial zirconia-containing PMMA bone cement.

A non-leaching antibacterial bone cement has been developed and evaluated. Chlorine- and bromine-containing furanone derivatives were synthesized and covalently coated onto the surface of zirconia filler particles, followed by mixing into a conventional poly(methyl methacrylate) bone cement. Flexural strength and bacterial viability were used to evaluate the modified cements. Effects of coated antibacterial moiety content, coated zirconia loading and halogen on furanone were investigated. Results showed that the experimental cement showed significant enhanced antibacterial function against bone-associated Gram-positive Staphylococcus aureus as well as Gram-negative Pseudomonas aeruginosa, as compared to commercial PMMA cement. The cement also exhibited a comparable flexural strength to and 3-14% higher flexural modulus than commercial PMMA bone cement. Increasing antibacterial moiety content and filler loading significantly enhanced antibacterial activity. Increasing antibacterial moiety content slightly increased both flexural strength and modulus of the modified cement. Increasing filler loading slightly increased flexural strength up to 7% loading and then decreased. The bromine-containing furanone modified cement showed a higher antibacterial activity than its chlorine counterpart. Antibacterial agent leaching tests exhibited that the modified experimental cement showed no leachable antibacterial components to surroundings. Within the limitations of this study, this experimental poly(methyl methacrylate) cement may find potential applications in orthopedics for reducing in-surgical and post-surgical infection after further investigations are conducted.