Bacterial Biofilm Growth Research Articles

Preparation, in vitro and in vivo assessment of novel carvacrol@pro-phytomicelles for the treatment of Salmonella enteritidis infection in mice

In livestock and poultry farming, the use of antibiotics has been abused, which seriously endangers human health. Thus, antibiotic alternatives are urgently needed. The phytochemical carvacrol (CAR) has attracted attention as an antibiotic alternative due to its excellent antibacterial activity and anti-inflammatory activity. However, CAR has high volatility and low water solubility, which seriously affect its antibacterial activity. In this study, two plant-derived small-molecule phytochemicals—glycyrrhizin and rebaudioside A—were selected as nanocarriers for the preparation of a novel solid pro-phytomicelle formulation named as CAR@PP. Using a simple fabrication method, the encapsulation efficiency of CAR reached 98.74 ± 1.14 %. CAR@PP was found to rapidly dissolve in water, resulting in a transparent solution (named as CAR@M) and a 59-fold increase in solubility compared to CAR. CAR@M contained uniform nanoparticles with a particle size, polydispersity index, and zeta potential of 3.52 ± 0.93 nm, 0.17 ± 0.01, and −10.63 ± 0.45 mV, respectively. The in vitro antibacterial activity of CAR@M was evaluated, and the minimum inhibitory concentration for the tested strains was 125–250 μg/ml. The antibacterial mechanisms were found that CAR@M disrupted the bacterial wall and biomembranes and efficiently inhibited bacterial biofilm growth. To the in vivo activity evaluation, treatment with 50 mg/kg CAR@M could effectively improve bacterial liver abscesses, decrease the inflammatory cytokine levels in the liver and cecum, and reduce the bacterial load in the liver and feces in Salmonella enteritidis-infected mice. In conclusion, CAR@PP is a promising alternative to antibiotics in livestock and poultry farming warranting further research.

Unveiling the Self-assembly and Therapeutic Efficacy of Antimicrobial Peptides SA4 Against Multidrug-Resistant A. baumannii.

Infections linked to Acinetobacter baumannii are one of the main risks of modern medicine. Biofilms formed by A. baumannii due to a protective extracellular polysaccharide matrix make them highly tolerant to conventional antibiotics and raise the possibility of antibiotic resistance. Antimicrobial peptides (AMPs) are gaining popularity due to their broad-spectrum actions and key properties of peptide self-assembly, making them a promising alternative to antibiotics. Here, we demonstrate that 12-residue synthetic self-assembled peptide SA4 nanostructures have enough antibacterial action to prevent the growth of mature bacterial biofilms. The SA4 peptide was successfully synthesized by using the solid-phase peptide synthesis method, and its self-assembly was prepared in water. The self-assembled peptide hydrogel formed nanotube structure was observed under a scanning electron microscope and further characterized to confirm their physical and molecular properties. The resulting hydrogel exhibits significant antibacterial activity against MDR A. baumannii strains (MDR-1 and MDR-2), responsible for many nosocomial infections. In addition, at various gel concentrations, this hydrogel has the potential to inhibit about 30-80% of biofilms formed by MDR strains. Furthermore, under a microscope, it has been observed that the rupture of the bacterial cell membrane and cell wall of A. baumannii cells is caused by peptide nanotubes generated by self-assemblies. Thus, peptide-based nanotubes present intriguing avenues for various biomedical applications. This is the first report of bacterial biofilm removal with SA4 peptide nanotubes, and offering a unique treatment for infections linked to biofilms.

Screening of Lithium Substituted Ag-TiO2 Nanoparticle Coating for Antibiofilm Application.

Bacterial infections and biofilm growth are common mishaps associated with medical devices, and they contribute significantly to ill health and mortality. Removal of bacterial deposition from these devices is a major challenge, resulting in an immediate necessity for developing antibacterial coatings on the surfaces of medical implants. In this context, we developed an innovative coating strategy that can operate at low temperatures (80 °C) and preserve the devices' integrity and functionality. An innovative Ag-TiO2 based coating was developed by ion exchange between silver nitrate (AgNO3) and lithium titanate (Li4Ti5O12) on glass substrates for different periods, ranging from 10 to 60 min. The differently coated samples were tested for their antibacterial and antibiofilm efficacy.

Effect of pH on hydroxyapatite coatings obtained by spray pyrolysis and their use as matrices for antibiotic adsorption by spin coating and release properties.

Calcium phosphate materials, particularly hydroxyapatite (HA), are extensively used in biomedical applications because of their prominence as primary inorganic constituents of human hard tissues. This study investigates the synthesis of HA coatings via spray pyrolysis using various precursors, including HA derived from bovine bone. The effects of pH on the formation and properties of HA coatings were systematically examined. Samples exposed to acidic conditions or left without pH adjustment led to the formation of HA, contrasting with the outcomes observed through dissolution methods. Different characterization techniques, such as scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray diffraction (XRD), were employed to evaluate the quality and crystallinity of the coatings. Among the samples, those exhibiting superior crystallinity and nanostructured features, including bovine HA, were selected for further surface functionalization with the antibiotic enrofloxacin using spin coating. As expected, the antibiotic loading on each material's surface depended on the amount of HA deposited on the substrate. However, the desorption results indicated that, in all cases, desorption persisted beyond 38 h, implying that HA-loaded matrices could be effective systems for controlled and prolonged drug release, which could be useful in dental or orthopedic implants for inhibiting the growth of bacterial biofilms.

Casein phosphopeptide/antimicrobial peptide co-modified SrTiO3 nanotubes for prevention of bacterial infections and repair of bone defects

Endowing titanium surfaces with multifunctional properties can reduce implant-related infections and enhance osseointegration. In this study, titanium dioxide nanotubes with strontium doping (STN) were first created on the titanium surface using anodic oxidation and hydrothermal synthesis techniques. Next, casein phosphopeptide (CCP) and an antimicrobial peptide (HHC36) were loaded into the STN with the aid of vacuum physical adsorption (STN-CP-H), giving the titanium surface a dual function of "antimicrobial-osteogenic". The surface of STN-CP-H has a suitable roughness and good hydrophilicity, which is conducive to osteoblasts. STN-CP-H had a 99 % antibacterial rate against S. aureus and E. coli and effectively prevented the growth of bacterial biofilm. Meanwhile, the antibacterial mechanism of STN-CP-H was initially explored with the help of transcriptome sequencing technology. STN-CP-H could greatly increase osteoblast adhesion, proliferation, and expression of osteogenic markers (alkaline phosphatase, runt-related transcription) when CCP and Sr worked together synergistically. In vivo, the STN-CP-H coating could effectively promote new osteogenesis around titanium implant bone and had no toxic effects on heart, liver, spleen, lung and kidney tissues. A potential anti-infection bone healing material, STN-CP-H bifunctional coating developed in this work efficiently inhibited bacterial infection of titanium implants and encouraged early osseointegration.

Bioelectronic Delivery of Potassium Ions Controls Membrane Voltage and Growth Dynamics in Bacteria Biofilms

AbstractBioelectrical signaling, or bioelectricity, is crucial in regulating cellular behavior in biological systems. This signaling, involving ion fluxes and changes in membrane potential (Vmem), is particularly important in the growth of bacterial biofilm. Current microfluidic-based methods for studying bacterial colonies are limited in achieving spatiotemporal control over ionic fluxes due to constant flow within the system. To address this limitation, we have developed a platform that integrates biofilm colonies with bioelectronic ion pumps that enable delivery of potassium (K+) ions, allowing for controlled manipulation of local potassium concentration. Our study examines the impact of controlled K+ delivery on bacterial biofilm growth patterns and dynamics. We observed significant changes in Vmem and coordination within the biofilms. Furthermore, we show that localized K + delivery is highly effective in controlling biofilm expansion in a spatially targeted manner. These findings offer insights into the mechanisms underlying bacterial signaling and growth, and suggest potential applications in bioengineering, synthetic biology, and regenerative medicine, where precise control over cellular signaling and subsequent tissue growth is required.

Bacterial biofilm formation in seafood: Mechanisms and inhibition through novel non‐thermal techniques

AbstractSeafoods are susceptible to microbial contamination due to their high moisture, nutrient contents and neutral pH. Among various microorganisms, biofilm‐forming bacteria pose a severe threat to the seafood supply chain as well as consumer health. Bacterial biofilm formation in seafood is primarily caused by Aeromonas hydrophila, Vibrio cholerae, V. vulnificus, V. parahaemolyticus, Salmonella spp., and Listeria monocytogenes. Biofilm formation is an important protective mechanism of microorganisms causing spoilage of seafood and disease threats to consumers. The attachment of microbes on the surface of seafood followed by the growth and proliferation of bacterial cells leads to the production of exopolymer compounds and the formation of biofilm. This biofilm is difficult to destroy or inhibit through conventional prevention/destruction techniques. The occurrence of bacterial strains/biofilms with more resistance to different preventive strategies is a big challenge for the seafood processing industry. This review covers the mechanisms of biofilm formation by bacteria and various non‐thermal processing approaches to prevent microbial contamination and biofilm formation in seafood products. The aforementioned non‐thermal processing techniques for the destruction of biofilm and quality control of seafood products include cold plasma treatment, irradiation, pulsed electric field technology, hydrostatic pressure processing, photosensitisation, natural bioactive compounds and so on. All these techniques effectively inhibit the bacterial biofilm and microbial growth without altering sensorial properties. However, further research validation and applications at the industry level are still required.

Sonochemical-assisted method for efficient synthesis of Cu-MOF and evaluating its antibacterial properties

Sonochemical-assisted method was used to synthesize copper metal-organic frameworks (Cu-MOF) nanostructures. The final products were examined by related techniques such as XRD patterns, SEM image, BET N2 adsorption/desorption technique and FTIR spectrum. Microtiter plates microbiological assay were used to investigate antibacterial properties and the results were analyzed using ANOVA and Tukey HSD tests. The results showed that Cu-MOF nanostructures have a mesoporous nature with an average particle size distribution around 60 nm. The final product had the property of preventing the growth of all tested bacteria in certain concentrations. Minimum Inhibitory Concentration (MIC) values were observed in the range of 30–100 ppm. It was also discovered that this nanostructure can not kill bacteria completely. In addition, the minimal inhibitory concentration for biofilm growth (MIC-B) of the nanostructure was investigated. The MIC-B analyzes demonstrated that the growth of bacterial biofilm decreased with increasing Cu-MOF concentration.

The Infected Polypropylene Mesh: When Does Biofilm Form and Which Antiseptic Solution Most Effectively Removes It?

BackgroundPolypropylene (PPE) mesh is commonly utilized to reconstruct catastrophic extensor mechanism disruptions in revision total knee arthroplasty. Unfortunately, these procedures are associated with a high rate of periprosthetic joint infection. The purpose of the current study was to: 1) visualize and quantify the progression of bacterial biofilm growth on PPE-mesh; and 2) determine which antiseptic solutions effectively remove viable bacteria. MethodsKnitted PPE mesh samples were cultured with either methicillin-sensitive Staphylococcus aureus (MSSA) or Escherichia coli (E. coli) for 7 days, with regular quantification of colony forming units (CFUs) and visualization using scanning electron microscopy to identify maturity. Immature (24 hour) and mature (72 hour) biofilm was treated with one of 5 commercial antiseptics for 3 minutes. A 0.05% chlorhexidine gluconate, a surfactant-based formulation of ethanol, acetic acid, sodium acetate, benzalkonium chloride, diluted povidone-iodine (0.35%), undiluted (10%) povidone-iodine, and 1:1 combination of 10% povidone-iodine and 3% hydrogen peroxide. A 3-log reduction in CFUs compared to saline was considered clinically meaningful. ResultsThe CFU counts plateaued, indicating maturity, at 72 hours for both MSSA and E. coli. The scanning electron microscopy confirmed confluent biofilm formation after 72 hours. The 10% povidone-iodine was clinically effective against all MSSA biofilms and immature E. coli biofilms. The 10% povidone-iodine with hydrogen peroxide was effective in all conditions. Only 10% povidone iodine formulations produced significantly (P < .0083) reduced CFU counts against mature biofilms. ConclusionsBacteria rapidly form biofilm on PPE mesh. Mesh contamination can be catastrophic, and clinicians should consider utilizing an antiseptic solution at the conclusion of mesh implantation. Undiluted povidone-iodine with hydrogen peroxide should be considered when attempting to salvage infected PPE mesh.

Bacterial biofilm growth and perturbation by serine protease from Bacillus sp.

In nature, bacteria are ubiquitous and can be categorized as beneficial or harmless to humans, but most bacteria have one thing in common which is their ability to produce biofilm. Biofilm is encased within an extracellular polymeric substance (EPS) which provides resistance against antimicrobial agents. Protease enzymes have the potential to degrade or promote the growth of bacterial biofilms. In this study, the effects of a recombinant intracellular serine protease from Bacillus sp. (SPB) on biofilms from Staphylococcus aureus, Acinetobacter baumannii, and Pseudomonas aeruginosa were analyzed. SPB was purified using HisTrap HP column and concentrated using Amicon 30 ultra-centrifugal filter. SPB was added with varying enzyme activity and assay incubation period after biofilms were formed in 96-well plates. SPB was observed to have contrasting effects on different bacterial biofilms, where biofilm degradations were observed for both 7-day-old A. baumannii (37.26%) and S. aureus (71.51%) biofilms. Meanwhile, SPB promoted growth of P. aeruginosa biofilm up to 176.32%. Compatibility between protein components in S. aureus biofilm with SPB as well as a simpler membrane structure morphology led to higher biofilm degradation for S. aureus compared to A. baumannii. However, SPB promoted growth of P. aeruginosa biofilm due likely to its degrading protein factors that are responsible for biofilm detachment and dispersion, thus resulting in more multi-layered biofilm formation. Commercial protease Savinase which was used as a comparison showed degradation for all three bacterial biofilms. The results obtained are unique and will expand our understanding on the effects that bacterial proteases have toward biofilms.

Photodynamic patterning of living bacterial biofilms with high resolutions for information encryption and antibiotic screening

AbstractControlling the growth of bacterial biofilms in a specific pattern greatly enhances the study of cell‐to‐cell interactions and paves the way for expanding their biological applications. However, the development of simple, cost‐effective, and highly resolved biopatterning approaches remains a persistent challenge. Herein, a pioneering photodynamic biopatterning technique for the creation of living bacterial biofilms with customized geometries at high resolutions is presented. First of all, an outstanding aggregation‐induced emission photosensitizer is synthesized to enable efficient photodynamic bacterial killing at a low concentration. By combining with custom‐designed photomasks featuring both opaque and transparent patterns, the viability of photosensitizer‐coated bacteria is successfully manipulated by controlling the degree of light transmittance. This process leads to the formation of living bacterial biofilms with specific patterns replicated from the photomask. Such an innovative strategy can be employed to generate living bacterial biofilms composed of either mono‐ or multispecies, with a spatial resolution of approximately 24 µm. Furthermore, its potential applications in information storage/encryption and antibiotic screening are explored. This study provides an alternative way to understand and investigate the intricate interactions among bacteria within 3D biofilms, holding great promise in the controlled fabrication of dynamic biological systems for advanced applications.

The dynamics of bacterial proliferation, viability, and extracellular polymeric substances in oral biofilm development

ObjectivesThis study investigated the relationship between bacterial growth, viability, and extracellular polymeric substances (EPS) formation in biofilms, particularly regarding resistance development. It also examined the impact of chemical factors on the EPS matrix and bacterial proliferation in oral biofilms. MethodsThree multi-species oral biofilms were incubated in anaerobic conditions. Three strains of Enterococcus faecalis were incubated in aerobic conditions. The incubation periods ranged from 0 h to 7 days for short-term biofilms, and from 3 to 90 days for long-term biofilms. Fluorescent labeling with carboxyfluorescein diacetate succinimidyl ester (CFSE) and flow cytometry were used to track EPS and bacterial growth. Confocal laser scanning microscopy (CLSM) assessed bacterial viability and EPS structure. Biofilms aged 7, 14, and 21 days were treated with 2 % chlorhexidine (CHX) and 1 % sodium hypochlorite (NaOCl) to evaluate their effects on EPS and bacterial proliferation. ResultsShort-term biofilms showed rapid bacterial proliferation and a gradual increase in EPS, maintaining stable viability. In the first two weeks, a significant rise in CFSE indicated growing maturity. From 14 to 90 days, EPS and CFSE levels stabilized. Following treatment, CHX significantly reduced bacterial proliferation, while NaOCl decreased EPS volume. ConclusionsBiofilm development involves a balance between bacterial proliferation and EPS production. The complexity of this process poses challenges in treating biofilm-associated infections, requiring strategies tailored to the biofilm's developmental stage. Clinical significanceFor effective root canal treatment, it is imperative to focus on reducing bacterial proliferation during the early stages of oral infections. In contrast, strategies aimed at minimizing EPS production could be more beneficial for long-term management of these conditions.

Dual roles of the conditional extracellular vesicles derived from Pseudomonas aeruginosa biofilms: Promoting and inhibiting bacterial biofilm growth

Antibiotic-resistant biofilm infections have emerged as public health concerns because of their enhanced tolerance to high-dose antibiotic treatments. The biofilm life cycle involves multiple developmental stages, which are tightly regulated by active cell-cell communication via specific extracellular signal messengers such as extracellular vesicles. This study was aimed at exploring the roles of extracellular vesicles secreted by Pseudomonas aeruginosa at different developmental stages in controlling biofilm growth. Our results show that extracellular vesicles secreted by P. aeruginosa biofilms during their exponential growth phase (G-EVs) enhance biofilm growth. In contrast, extracellular vesicles secreted by P. aeruginosa biofilms during their death/survival phase (D-EVs) can effectively inhibit/eliminate P. aeruginosa PAO1 biofilms up to 4.8-log10 CFU/cm2. The inhibition effectiveness of D-EVs against P. aeruginosa biofilms grown for 96 h improved further in the presence of 10–50 μM Fe3+ ions. Proteomic analysis suggests the inhibition involves an iron-dependent ferroptosis mechanism. This study is the first to report the functional role of bacterial extracellular vesicles in bacterial growth, which depends on the developmental stage of the parent bacteria. The finding of D-EV-activated ferroptosis-based bacterial death may have significant implications for preventing antibiotic resistance in biofilms.

Submerged macrophyte promoted nitrogen removal function of biofilms in constructed wetland

Biofilm is one of the important factors affecting nitrogen removal in constructed wetlands (CWs). However, the impact of submerged macrophyte on nitrogen conversion of biofilms on leaf of submerged macrophyte and matrix remains poorly understood. In this study, the CWs with Vallisneria natans and with artificial plant were established to investigate the effects of submerged macrophyte on nitrogen conversion and the composition of nitrogen-converting bacteria in leaf and matrix biofilms under high ammonium nitrogen (NH4+-N) loading. The 16S rRNA sequencing method was employed to explore the changes in bacterial communities in biofilms in CWs. The results showed that average removal rates of total nitrogen and NH4+-N in CW with V. natans reached 71.38% and 82.08%, respectively, representing increases of 24.19% and 28.79% compared with the control with artificial plant. Scanning electron microscope images indicated that high NH4+-N damaged the leaf cells of V. natans, leading to the cellular content release and subsequent increases of aqueous total organic carbon. However, the specific surface area and carrier function of V. natans were unaffected within 25 days. As a natural source of organic matters, submerged macrophyte provided organic matters for bacterial growth in biofilms. Bacterial composition analysis revealed the predominance of phylum Proteobacteria in CW with V. natans. The numbers of nitrifiers and denitrifiers in leaf biofilms reached 1.66 × 105 cells/g and 1.05 × 107 cells/g, as well as 2.79 × 105 cells/g and 7.41 × 107 cells/g in matrix biofilms, respectively. Submerged macrophyte significantly increased the population of nitrogen-converting bacteria and enhanced the expressions of nitrification genes (amoA and hao) and denitrification genes (napA, nirS and nosZ) in both leaf and matrix biofilms. Therefore, our study emphasized the influence of submerged macrophyte on biofilm functions and provided a scientific basis for nitrogen removal of biofilms in CWs.

Penurunan Aktivitas Biofilm Strain MRSA 22156 oleh Tanaman Saga (Abrus precatorius L.)

The aim of this research is to determine the effect of using saga extract on bacterial biofilm growth, and to determine the most appropriate dose of saga extract in reducing MRSA 22156 biofilm activity. This research uses the Dowd method to determine the total flavonoid content of A. precatorius L leaves. Treatment reduction in MRSA 22156 biofilm growth using the Plate Biofilm Assay method with three repetitions. The results of the research show that total flavonoids can reduce the growth of MRSA 22156 biofilms. The use of total flavonoids from A. precatorius L. leaves in concentrations of 800 mgL-1 to 25 mgL-1 has been proven to have an effect on biofilm growth activity as indicated by the reduction in MRSA 22156 biofilm growth by 67.9% and 25%. The ethanol extract of A. precatorius L. has anti-biofilm activity in reducing the growth of Staphylococcus aureus bacteria that are resistant to Methicillin.

Targeted modulation and enhancement of macrophages via sonodynamic therapy-driven cupferroptosis-like stress for implant-associated biofilm infections

As the vanguard against implant-associated infections (IAIs), macrophages facilitate the phagocytosis and pro-inflammatory polarization of planktonic and biofilm infections, but indirectly serve as a “Trojan horse” for intracellular bacterial infections. Effective drug delivery and macrophage immunomodulation to eliminate intracellular infections will contribute to the treatment of refractory IAIs and prevention of relapse. Herein, we developed an ultrasound-responsive copper-based nanoparticle targeting macrophages by encapsulating copper in a porphyrin metal-organic framework and surface-grafting D-Mannosamine to synthesize CuTCPP@MOF nanodots@ Mannosamine (CTMM) with macrophage-targeting functionality. Concurrent generation of hydroxyl radicals and singlet oxygen in biofilm microenvironment mediated by ultrasound disrupts the biofilm structure, achieving CTMM overload and interfering with bacterial biofilm growth and metabolism. By regulating macrophage oxidative stress metabolism, CTMM induced a cupferroptosis-like stress, characterized by concurrent cuproptosis and ferroptosis, to eliminate intracellular infections. Transcriptomic analysis verified that the cupferroptosis-like stress based on GPX4 axis modulation further promotes macrophage inflammatory activation. This CTMM+US-mediated bacterial clearance in biofilms and macrophage immunomodulation informs the "Trojan War" strategy to combat IAIs clinically.

Traditional and biodegradable plastics host distinct and potentially more hazardous microbes when compared to both natural materials and planktonic community

Microplastic particles are persistent micropollutants that provide a substrate for the growth of bacterial biofilms, posing a threat to the environment. This study explores the changes in commercially available food containers made of conventional (polypropylene PP, polyethylene terephthalate PET), innovative biodegradable (Mater-Bi) and natural (wood and cellulose) materials, when introduced in the surface waters of Lake Maggiore for 43 days. Spectral changes revealed by FT-IR spectroscopy in PET and Mater-Bi, and changes in thermal properties of all human-made material tested indicated a degradation process occurred during environmental exposure. Despite similar bacterial richness, biofilms on PET, PP, and Mater-Bi differed from natural material biofilms and the planktonic community. Human-made material communities showed a higher proportion of potential pathogens, with PET and PP also exhibiting increased abundances of antibiotic resistance genes. Overall, these findings stress the need for dedicated strategies to curb the spread of human-made polymers in freshwaters, including innovative materials that, due to their biodegradable properties, might be perceived less hazardous for the environment.

A review of bacterial biofilm formation and growth: rheological characterization, techniques, and applications

A review of bacterial biofilm formation and growth: rheological characterization, techniques, and applications

Three-Dimensional Cellular Automaton for Modeling of Self-Similar Evolution in Biofilm-Forming Bacterial Populations

Bacterial populations often form colonies and structures in biofilm. The paper aims to design suitable algorithms to simulate self-similar evolution in this context, specifically by employing a hybrid model that includes a cellular automaton for the bacterial cells and their dynamics. This is combined with the diffusion of the nutrient (as a random walk), and the consumption of nutrients by biomass. Lastly, bacterial cells divide when reaching high levels. The algorithm computes the space-time distribution of biomass under limited nutrient conditions, taking into account the collective redistribution of nutrients. To achieve better geometry in this modified model approach, truncated octahedron cells are applied to design the lattice of the cellular automaton. This allows us to implement self-similar realistic bacterial biofilm growth due to an increased number of inner relations for each cell. The simulation system was developed using C# on the Unity platform for fast calculation. The software implementation was executed in combination with the procedure of surface roughness measurements based on computations of fractional dimensions. The results of the simulations qualitatively correspond to experimental observations of the population dynamics of biofilm-forming bacteria. Based on in silico experiments, quantitative dependencies of the geometrical complexity of the biofilm structure on the level of consumed nutrients and oxygen were revealed. Our findings suggest that the more complex structure with a fractal dimension of the biofilm boundaries (around 2.6) corresponds to a certain range of nutrient levels, after which the structure degenerates and the biofilm homogenizes, filling the available space provided and tending towards a strictly 3D structure. The developed hybrid approach allows realistic scenario modeling of the spatial evolution of biofilm-forming bacterial populations and specifies geometric characteristics of visualized self-similar biofilm bacterial structures.

Sucrose-mediated formation and adhesion strength of Streptococcus mutans biofilms on titanium

Biofilms consist of bacterial cells surrounded by a matrix of extracellular polymeric substance (EPS), which protects the colony from many countermeasures, including antibiotic treatments. Growth and formation of bacterial biofilms are affected by nutrients available in the environment. In the oral cavity, the presence of sucrose affects the growth of Streptococcus mutans that produce acids that erode enamel and form dental caries. Biofilm formation on dental implants commonly leads to severe infections and can restrict osseointegration necessary for the implant to be successful. This work determines the effect of sucrose concentration on biofilm EPS formation and adhesion of Streptococcus mutans, a common oral colonizer, to titanium substrates simulating common dental implants. Biofilm formation and profiles are visualized at high magnification with scanning electron microscopy (SEM). Large mounds and complex structures consisting of bacterial cells and EPS can be seen in biofilms at sucrose concentrations that are favorable for biofilm growth. The laser spallation technique is used to apply stress wave loading to the biofilm, causing the biofilm to delaminate at a critical tensile stress threshold. The critical tensile stress threshold is the adhesion strength. Because laser spallation applies the stress loading to the rear of the substrate, bulk adhesion properties of the biofilm can be determined despite the heterogenous composition and low cohesion strength of the biofilm. Statistical analysis reveals that adhesion strength of biofilms initially increase with increasing sucrose concentration and then decrease as sucrose concentration continues to increase. The adhesion strength of bacterial biofilms to the substrate in this study is compared to the adhesion of osteoblast-like cells to the same substrates published previously. When sucrose is present in the biofilm growth environment, S. mutans adhesion is higher than that of the osteoblast-like cells. Results of this study suggest sucrose-mediated S. mutans biofilms may outcompete osteoblasts in terms of adhesion during osseointegration, which could explain higher rates of peri-implant disease associated with high sugar diets. Further studies demonstrating adhesion differentials between biofilms and cells including co-cultures are needed and motivated by the present work.