Adhesion Of Bacteria Research Articles

Surface characteristics, cytotoxicity, and microbial adhesion of 3D-printed hybrid resin-ceramic materials for definitive restoration

ObjectiveThis study investigated the surface properties, cytotoxicity, and microbial adhesion of 3D-printed specimens made from hybrid resin-ceramic materials intended for use in definitive crowns. MethodsDisc-shaped specimens were 3D-printed using six different hybrid resin-ceramic materials recommended for definitive restorations: Crowntec (CT), VarseoSmile Crown Plus (VS), Tera Harz TC-80DP Graphy (TH), C&B Permanent ODS (CB), Formlabs Permanent Crown (FP), and HeyGears (HG). Surface topography, surface roughness, and water contact angle values were measured for each material (n = 6). Cytotoxicity was assessed using direct contact and extract tests on human gingival fibroblasts (n = 4). Additionally, the adhesion of mixed oral bacteria to the surfaces of the specimens was evaluated by counting colony-forming units (CFUs) after a 2-hour incubation period (n=6). ResultsThe TH group exhibited significantly lower surface roughness (Ra: 0.28 ± 0.13 μm) compared to the other materials (CT: 1.87 ± 0.34 μm; VS: 1.13 ± 0.09 μm; CB: 2.91 ± 0.27 μm; FP: 2.50 ± 0.08 μm; HG: 1.50 ± 0.55 μm). The VS group had the highest water contact angle (129.5 ± 1.1°), indicating greater hydrophobicity, in contrast to the other groups (CT: 72.6 ± 2.1°; TH: 75.0 ± 0.3°; CB: 69.1 ± 0.2°; FP: 93.0 ± 1.6°; HG: 77.7 ± 0.3°). Cytotoxicity testing showed no harmful effects, as relative cell viability exceeded 70%, and lactate dehydrogenase (LDH) release remained below 30% for all materials. The TH specimens also demonstrated the lowest bacterial adhesion. ConclusionsThe surface characteristics of the tested resin-ceramic materials varied significantly, with TH showing the smoothest surface and the least bacterial adhesion. All materials were found to be non-toxic. Therefore, TH material has the potential to provide definitive restorations with less microbial adhesion. Clinical SignificanceThe type of resin-ceramic material significantly affects the surface properties of 3D-printed specimens. These findings are crucial for selecting the appropriate resin-ceramic material for definitive restorations.

Dietary carbohydrates modulate Streptococcus mutans adherence and bacterial proteome.

Streptococcus mutans adherence to the tooth surface and subsequent biofilm development is modulated by the carbohydrate source, but the corresponding effect on bacterial proteome has not been previously studied. This study aimed to assess the effect of different carbohydrates on S. mutans viability and bacterial proteome at two-time points, early attachment (8 h) and biofilm maturation (24 h). Hydroxyapatite (HAp) discs coated with parotid saliva proteins were inoculated with S. mutans UA159 in Tryptone Soy Broth without dextrose supplemented with one of the following carbohydrates (n=12/treatment/time point): 1% Sucrose (S); 0.525% Glucose + 0.525% Fructose (G+F); 10% Xylitol (X); 10% Xylitol + 1% Sucrose (X+S); or culture medium without supplementation as negative control (C). Once inoculated, HAp discs were incubated for 8 h or 24 h at 37 °C and 10% CO2. After each incubation period, adhered bacteria were quantified using the plate-counting method for 6 HAp discs/group, and the remaining 6 HAp discs/group were used to extract bacterial cell wall proteins. Extracted proteins were analyzed using liquid chromatography coupled with mass spectrometry and then classified by their biological process. The study was conducted in three independent assays and the number of bacteria adhered to the HAp discs was determined at each time point and analyzed by two-way ANOVA followed by Bonferroni test (=5%). The results suggest that xylitol significantly repressed bacterial adherence and metabolism at 8 h and 24 h; however, bacterial adherence and metabolism were significantly enhanced when xylitol was combined with sucrose, showing no negative effect on S. mutans at both time points. Bacterial proteome was modulated by the carbohydrate source. The cariogenicity of S. mutans biofilms may be reduced by the alternative sweetener xylitol; however, the combination with fermentable sugars may inhibit such a beneficial effect.

Macrophage Immunomodulation and Suppression of Bacterial Growth by Polydimethylsiloxane Surface-Interrupted Microlines’ Topography Targeting Breast Implant Applications

Since breast cancer is one of the most common forms of cancer in women, silicone mammary implants have been extensively employed in numerous breast reconstruction procedures. However, despite the crucial role they play, their interaction with the host’s immune system and microbiome is poorly understood. Considering this, the present work investigates the immunomodulatory and bacterial mitigation potential of six textured surfaces, based on linear step-like features with various regular and irregular multiscaled arrangements, in comparison to a flat PDMS surface. We hypothesise that the chosen surface geometries are capable of modulating the cellular response through mechanical interdigitation within the multiscaled surface morphology, independent of the surface chemical properties. Each type of sample was characterised from a physico-chemical and biological points of view and by comparison to the flat PDMS surface. The overall results proved that the presence of linear multiscaled step-like features on the PDMS surface influenced both the surface’s characteristics (e.g., surface energy, wettability, and roughness parameters), as well as the cellular response. Thus, the biological evaluation revealed that, to different degrees, biomaterial-induced macrophage activation can be mitigated by the newly designed microtextured surfaces. Moreover, the reduction in bacteria adherence up to 90%, suggested that the topographical altered surfaces are capable of suppressing bacterial colonisation, therefore demonstrating that in a surgical environment at risk of bacterial contamination, they can be better tolerated.

Zwitterionic Polymers for Biomedical Applications: Antimicrobial and Antifouling Strategies toward Implantable Medical Devices and Drug Delivery.

Poly(ethylene glycol) (PEG) is extensively utilized in biomedical applications due to its biocompatibility; however, its thermal instability and susceptibility to oxidative degradation significantly constrain its long-term effectiveness. Zwitterionic polymers, characterized by their distinctive structure, enhanced stability, and superior biocompatibility, offer a more advantageous alternative. These polymers exhibit super hydrophilicity, resist nonspecific protein adsorption, and maintain stability in biological environments due to their charge-neutral ionic nature. Zwitterionic polymers enhance anticancer drug delivery by precisely targeting tumor cells and facilitating an efficient drug release. Their inherent antifouling properties and prolonged circulation within the bloodstream render them highly suitable for redox-sensitive drug carriers, thereby augmenting the antitumor efficacy. Moreover, zwitterionic polymers markedly mitigate biofouling in implants, biosensors, and wound dressings, thereby improving both their functionality and their therapeutic outcomes. These advantages arise from the formation of robust hydration layers, which significantly enhance the hemocompatibility and inhibit the adhesion of proteins, platelets, and bacteria. Zwitterionic polymers, including sulfobetaine (SB), phosphorylcholine (PC), and carboxybetaine (CB), are increasingly employed in blood-contacting devices and as effective coating materials for implantable devices. This mini-review paper aims to explore the recent diverse biomedical applications of zwitterionic polymers and highlight their advantageous properties compared with unmodified polymers. We will cover their use in drug delivery systems, tumor targeting nanocarriers, antibiofouling and antibacterial activities in implantable devices, tissue engineering, and diagnostic devices, demonstrating how their unique properties can translate into different applications. Through this exploration, this Perspective will display the potential of zwitterionic polymers as innovative polymer materials in the field of biomedical engineering and beyond.

The outer membrane protein, OMP71, of Riemerella anatipestifer, mediates adhesion and virulence by binding to CD46 in ducks

The Riemerella anatipestifer bacterium is known to cause infectious serositis in ducklings. Moreover, its adherence to the host’s respiratory mucosa is a critical step in pathogenesis. Membrane cofactor protein (MCP; CD46) is a complement regulatory factor on the surface of eukaryotic cell membranes. Bacteria have been found to bind to this protein on host cells. Outer membrane proteins (OMPs) are necessary for adhesion, colonisation, and pathogenicity of Gram-negative bacteria; however, the mechanism by which R. anatipestifer adheres to duck cells remains unclear. In this study, pull-down assays and LC–MS/MS identified eleven OMPs interacting with duck CD46 (dCD46), with OMP71 exhibiting the strongest binding. The ability of an omp71 gene deletion strain to bind dCD46 is weaker than that of the wild-type strain, suggesting that this interaction is important. Further evidence of this interaction was obtained by synthesising OMP71 using an Escherichia coli recombinant protein expression system. Adhesion and invasion assays and protein and antibody blocking assays confirmed that OMP71 promoted the R. anatipestifer YM strain (RA-YM) adhesion to duck embryo fibroblasts (DEFs) by binding to CD46. Tests of the pathogenicity of a Δomp71 mutant strain of RA-YM on ducks compared to the wild-type parent supported the hypothesis that OMP71 was a key virulence factor of RA-YM. In summary, the finding that R. anatipestifer exploits CD46 to bind to host cells via OMP71 increases our understanding of the molecular mechanism of R. anatipestifer invasion. The finding suggests potential targets for preventing and treating diseases related to R. anatipestifer infection.

Studies on the Probiotic, Adhesion, and Induction Properties of Artisanal Lactic Acid Bacteria: to Customize a Gastrointestinal Niche to Trigger Anti-obesity Functions.

The primary goals of this work are to explore the potential of probiotic lactic acid bacteria's (LAB) mucin/mucus layer thickening properties and to identify anti-obesity candidate strains that improve appropriate habitat for use with the Akkermansia group population in the future. The HT-29 cell binding, antimicrobial properties, adhesion to the mucin/mucus layer, growth in the presence of mucin, stability during in vitro gastrointestinal (GI) conditions, biofilm formation, and mucin/mucus thickness increment abilities were all assessed for artisanal LAB strains. Sixteen LAB strains out of 40 were chosen for further analysis based on their ability to withstand GI conditions. Thirteen strains remained viable in simulated intestinal fluid, while most showed high viability in gastric juice simulation. Furthermore, 35.9-65.4% of those 16 bacteria adhered to the mucin layer. Besides, different lactate levels were produced, and Streptococcus thermophilus UIN9 exhibited the highest biofilm development. In the HT-29 cell culture, the highest mucin levels were 333.87µg/mL with O. AK8 at 50mM lactate, 313.38µg/mL with Lactobacillus acidophilus NRRL-B 1910 with initial mucin, and 311.41µg/mL with Lacticaseibacillus casei NRRL-B 441 with initial mucin and 50mM lactate. Nine LAB strains have been proposed as anti-obesity candidates, with olive isolates of Lactiplantibacillus plantarum being particularly important due to their ability to avoid mucin sugar consumption. Probiotic LAB's attachment to the colonic mucosa and its ability to stimulate HT-29 cells to secrete mucus are critical mechanisms that may support the development of Akkermansia.

Effect of Egg Washing and Hen Age on Cuticle Quality and Bacterial Adherence in Table Eggs

The cuticle covering the outer surface of an eggshell functions as both a physical and chemical barrier against invading microorganisms. Contamination of eggs by microbial pathogens progresses in four stages: bacterial attachment to the egg surface, penetration through the cuticle and eggshell, multiplication within the underlying membranes, and the final stage of contaminating the egg contents. Therefore, it is important to study bacterial count at the first point of contact, i.e., on the surface of the eggs. In this study, we have evaluated the impact of differences in cuticle quality (due to egg washing and hen age) on bacterial load. We compared bacterial adherence on the eggshell surface of white eggs which were either washed (graded) or unwashed (ungraded), collected from Lohmann laying hens of different ages: early (24–28 weeks), mid-lay (44–48 weeks), and late (66–70 weeks). We aimed to determine the impact of hen age and egg washing on differences in cuticle quality and bacterial adherence. Our results indicate that hen age (up to 70 weeks) and commercial egg washing do not significantly impact bacterial adherence on eggshell surfaces. We have developed a novel method using green fluorescent protein (GFP)-expressing Salmonella typhimurium to estimate adherence of bacteria to the eggshell surface, with independent measurement of autofluorescence to quantitate cuticle deposition. S. typhimurium were localized, adhering to cracks visible on the outer cuticle in ungraded eggs, indicating that egg-associated pathogens usually enter the egg interior either through respiratory pores in eggshells or through shell micro-cracks. The results of this study can be utilized to optimize innovative methods for predictive microbiology in order to achieve egg safety.

Inhibition of bacteria adhesion and biofilm formation using a precisely structured nitric oxide-releasing coating with repeatedly renewing antimicrobial and antifouling ability

Bacterial adhesion and colonization usually causes the formation of biofilm that cannot be easily eradicated by common antibiotics and disinfectants. Surface covalent immobilization of nitric oxide (NO)-releasing polymer is an effective surface modification strategy to combat the threat from bacteria. Although surface covalent strategy avoided the leaching of toxic NO donors, the existence of hydrophobic NO donors on topsurface could inevitably accelerate the microorganism adhesion and accumulation, and the NO-releasing period of time was still very limited. In this work, we prepared an antimicrobial and antibiofilm surface via tethering a precision-structured NO-releasing copolymer brush to an organic-inorganic hybrid hard coating (> 5H pencil hardness) on one end. The precision-structured diblock copolymer brush was composed of a surface antifouling block and a subsurface antimicrobial block of NO-releasing units. A typical effect of “kill two birds with one stone” was observed, the NO releasing subsurface segment within a polymeric matrix prolongs NO release while also preventing surface fouling caused by hydrophobic NO donors on top surface. The impacts of polymer architecture on bioactivity was detailed investigated, and the block copolymer brush exhibited the optimal inhibitory effects against bacteria colonization and biofilm formation. The developed bioactive diblock copolymer brush was grafted from a hard hybrid persistent coating that embeds considerable initiation sites for surface modification. Thanks to the presence of initiator throughout the hybrid network, the initiation layer could initiate polymerization repeatedly after the polymer brushes are worn off at high pressure. Thus, the coating exhibited repeatedly renewing antimicrobial and antifouling ability after multiple abrasion cycles. This work not only developed a novel strategy for regulating NO releasing via modulating the polymer architecture, but provided a feasible route for obtaining a robust initiator layer for synthesizing polymer brush for antimicrobial and antifouling applications.

Bactericidal and Antifouling Coatings with the "Killing-Repelling-Killing" Triple Function Based on Cationic Copolymers with Structure Conversion and Capsaicin Analogue Release.

The capsaicin analogue N-(4-hydroxy-3-methoxybenzyl) acrylamide (HMBA) was linked with polylauryl methacrylate-b-poly(2-(N,N-dimethylamino)ethyl methacrylate) (PLMA-b-PDMAEMA) via a quaternization reaction with 4-(acrylamidomethyl)-2-methoxyphenyl 2-chloroacetate (AAMPCA). The amphiphilic copolymers were capable of transforming its structure in response to the solvent change from aprotic to protic, which was verified by the 1H NMR spectrum. The resulting cationic copolymers underwent a hydrolysis process in water, yielding zwitterionic groups on surfaces. Meanwhile, the bactericidal reagent HMBA was released. It was proved that the hydrolysis rate of the copolymers accelerated with higher temperature, higher pH value, and higher hydrophilic block units. And the controllable, sustainable release of HMBA was achieved with copolymer-mediated hydrolysis. Protein-repellent and bactericidal tests on the surface of the coating proved that antifouling and bactericidal performances of the coating correlated to the structure conversion abilities of the corresponding copolymer. The dynamic monitoring of Escherichia coli adhesion in 3 h evidenced the antifouling and bactericidal process of copolymers with different block ratios and concentrations. The coating incorporated with 3% PLMA120-b-(PDMAEMA-AAMPCA)120 in polylactic acid base materials showed an adhesion ratio of E. coli less than 1% within 1 h, and the survival ratio of the adhered bacteria is <1%, suggesting its rapid speed and high efficiency in "bacterial repelling and killing". Also, the PLMA120-b-(PDMAEMA-AAMPCA)120 copolymer demonstrated enhanced bactericidal ability compared with the mixture of cationic poly(laruyl methacrylate)120-b-poly(carboxybetaine methacrylate ester)120 (PLMA120-b-PCBMAE120) and free HMBA. The lowest minimal inhibitory concentration was 0.078 mg/mL against Staphylococcus aureus and 0.312 mg/mL against E. coli, respectively.

Comparative Adhesion of Pseudomonas aeruginosa to Human Oral Mucosal Epithelial Cells and Polystyrene Surfaces

Background: The adhesion of bacteria to biotic and abiotic surfaces reflects their ability to cause infectious diseases. The distinction between the ability of Pseudomonas aeruginosa bacteria to adhere to biotic and abiotic surfaces is not clear in the literature. Objectives: The current study aims to shed light on the extent of similarities and differences between P. aeruginosa in terms of their ability to adhere to different biotic and abiotic surfaces. Materials and Methods: ten isolates of P. aeruginosa were isolated from 100 wound and burn swabs. The isolates were identified using biochemical and phenotypic tests in addition to VITIK-II technology. The susceptibility to antibiotics was estimated using the disc diffusion method. The microtiter-spectrophotometric was used to measure the biofilm formation on polystyrene microtiter plates. The Human oral mucosal epithelial (OMECs) were used to evaluate the adhesion of P. aeruginosa isolates. Plate count and direct bacterial count were used to count the adhered bacteria to human OMECs in vitro. Results: Norfloxacin showed the highest antibacterial effect while the lowest antibacterial effect was seen in the case of using when Amoxicillin and Cefixime (all isolates were resistant). All isolates from, Pa2 form the highest biofilm followed by Pa6, while the lowest biofilm formation was seen in the case of Pa4. While Pa 6 showed the highest ability to attach to human OMECs followed by Pa2. Conclusion: It can be concluded that P. aeruginosa isolates are resistant to most antibiotics and that their ability to adhere and biofilm formation depends on the type of surface.

Polydopamine Nanoconjugate‐Coated Masks: An Efficient Barrier Against Airborne Pathogens

AbstractCurrent global burden of respiratory tract infections (RTIs) has thrown new challenges for researchers to design and develop efficient and multifaceted face masks. Airborne pathogens are highly transmissible through droplet nuclei/aerosol (&lt;5 μm) which can be significantly reduced by wearing masks. Currently available disposable surgical masks have disparate limitations in terms of surface wettability, microbial penetration, dermal infections due to anaerobic reactions beneath the surface, discomfort, and eruption of chronic obstructive pulmonary diseases (COPD). These mask surfaces get contaminated with airborne pathogens along with commensal bacterial strains. Generation of high temperature and humidity during repetitive breathing allow bacteria to penetrate from the masks to respiratory tract causing serious complications. Till date, functionalized face masks with different microbicidal agents including nanomaterials experience restricted leaching of antimicrobial agents, cytotoxic effects, short‐lived microbicidal effects, inflammatory responses, negative impact on ecosystem, etc. To address these limitations, here, we have attempted fabrication of mussel mimetic biopolymer (PDA) and its aminoglycoside‐nanoconjugate‐coated face masks. Formation of a uniform layer on the surface not only acted as a potential barrier for infectious pathogens but also inhibited their entry, deactivated the adhered bacteria, imparted non‐immunogenic response with no harmful effects on the wearer's skin owing to the biocompatible nature and pH compatibility of PDA and their conjugates. Amongst all, PDA‐kanamycin (PDA−K)‐coated masks exhibited excellent microbicidal activity with the least bacterial infiltration efficiency.

Robust, intelligent photochromic cotton fabrics with superamphiphobicity and expedient UV detection ability

Multifunctional photochromic cotton fabrics have enormous application potential in our daily lives, but still suffer from poor durability, slow coloration, tedious fabrication process, and short service life. The hydrophilic and polysaccharide characteristics of cotton fabrics make them vulnerable to bacteria adhesion and proliferation. Herein, intelligent photochromic cotton fabrics featured with durable superamphiphobicity are fabricated by in situ growth of ZIF-8 nanoparticles encapsulating spirooxazine (SP) photochromic dyes on the fabric surface, followed by low surface energy treatment using a fluorocarbon resin (FR) via a dip-coating method. The resultant SP@ZIF-8/FR cotton fabrics exhibit superamphiphobicity with contact angles of over 150° to both water and oils (surface tension ≥30 mN/m). When exposed to UV light, the fabric rapidly changes its color from white to blue within 10 s and fades in 2 min under visible room light. The developed SP@ZIF-8/FR coating is durable against numerous damages in daily usage, such as machine laundry, abrasion and UV irradiation, without losing photochromic and superamphiphobic properties. The high durability and excellent reversible color response allow the coated fabric to intuitively and quantitatively monitor outdoor UV intensity. Owing to the presence of ZIF-8, the treated fabrics also show excellent antibacterial property with antibacterial rates of over 99 % against both E. coli and S. aureus. The developed fabric coating can be engineered to visually assess the UV intensity in the outdoor environment.

Preventing bacterial adhesion to skin by altering their physicochemical cell surface properties specifically

The adhesion of bacteria to surfaces is associated with physicochemical and biological interactions. The present investigations provide new results about the differential adhesion levels of skin bacteria using a representative 3D skin model which mainly relies on the different physicochemical properties of the respective surfaces. Modulation of the adhesion of bacteria and thus their colonization, may occur by adjusting the physicochemical properties of the epidermal and bacterial surfaces. Lewis acid and hydrophobicity were the most strongly correlated parameters with the antiadhesion properties of the tested compounds. Modulation of physicochemical properties appears to be the primary driver of reduced Staphylococcus aureus adhesion in this study, with no significant changes observed in the expression of genes associated with classical adhesion pathways.

Biofilm Production Activity and Antibiotics Susceptibility of Streptococcus mutans Isolates from Patients with Dental Caries

Dental caries, a prevalent chronic infectious ailment, commences with the adherence of cariogenic bacteria to the teeth, particularly Streptococcus mutans. These microorganisms utilize sugars to produce acid, resulting in the gradual demineralization of the dental structure. Streptococcus mutans a gram positive bacterium is found to play the most significant role in the dental caries in terms of being the earliest colonizer in the formation of caries. The current research focuses on the isolation and identification of the Streptococcus mutans pathogen in individuals affected by dental caries. The isolated strains were cultured on specific media, with one hundred samples of dental caries obtained from human teeth. Among these samples, fifty were identified as positive bacterial isolates utilizing MS-agar (Mitist Salivares agar), with ten isolates categorized under the group Streptococci and twenty isolates identified as S. mutans. Identification of bacterial isolates was carried out based on morphological and cultural characteristics, biochemical tests, and VITEK-II. Additionally, an antibiogram assay was conducted to assess the susceptibility and resistance of the pathogen to various drugs. The findings revealed that all locally isolated S. mutans strains displayed 100% resistance to Cefepime, while showing different levels of resistance and susceptibility to other antibiotics such as Doxycycline, Amoxicillin-clavulanic acid, Trimethoprim-sulfamethoxazole, Vancomycin, Tetracycline, Ampicillin, and Meropenem. In this study, ELISA was employed to differentiate between Streptococcus. mutans isolates that produced biofilms and those that did not by utilizing the optical density at 630 nm. Bacterial biofilms contribute to chronic diseases that pose challenges in treatment. Twenty distinct isolates were evaluated for their biofilm-forming capabilities, with 9 (45%) identified as strong biofilm producers, 7 (35%) classified as moderate biofilm producers, and 4 (20%) categorized as weak biofilm producers

Bacterial adhesion and surface roughness of particulate-filled and short fiber-reinforced composites.

The objective of the study was to assess the initial adhesion of Streptococcus mutans (S. mutans) and surface roughness of different particulate-filled (PFC) and short fiber-reinforced (SFRC) composites. Five PFC composites (CeramX Universal, Filtek Universal, Omnichroma, Tetric Prime and Venus Diamond) and four SFRC composites (everX Posterior, everX Flow Bulk, everX Flow Dentin and experimental packable SFRC) were tested in this study. A non-contact 3D profilometer was employed to assess the surface roughness (Ra) of the polished specimens (using 4000-grit abrasive paper). For the bacterial adhesion test, the specimens (n = 5/group) were immersed in a solution of S. mutans to facilitate initial adhesion. To determine the number of cells on the surfaces of the discs as colony-forming units (CFU), the vials holding the microbial samples were highly agitated using a vortex machine. Subsequently, the samples were diluted multiple times and anaerobically incubated for 48h at 37°C on Mitis Salivarius Agar plates (Difco) supplemented with bacitracin. Bacterial adherence assessment was performed using SEM. The data were analyzed using ANOVA. All tested PFC and SFRC composites showed similar adhesion of S. mutan. The lowest Ra values (0.26µm) (p < 0.05) were found in the flowable SFRCs (everX Flow Bulk & Dentin), while the highest values (p < 0.05) were observed in CeramX and everX Posterior (0.42µm). Experimental SFRC had comparable Ra value (0.38µm) than other commercial composites. The presence of short microfibers in the composite appeared to have no adverse effects on the initial adhesion of bacteria or the surface roughness.

Inhibition of Oral Pathogenic Bacteria, Suppression of Bacterial Adhesion and Invasion on Human Squamous Carcinoma Cell Line (HSC-4 Cells), and Antioxidant Activity of Plant Extracts from Acanthaceae Family.

Medicinal plants have traditionally been used to treat various human diseases worldwide. In this study, we evaluated the leaf extracts of plants from the Acanthaceae family, specifically Clinacanthus nutans (Burm.f.) Lindau, Thunbergia laurifolia Lindl., and Acanthus ebracteatus Vahl., for their compounds and antioxidant activity. The ethanolic extracts of A. ebracteatus showed the highest total phenolic content at 22.55 mg GAE/g extract and the strongest antioxidant activities, with IC50 values of 0.24 mg/mL and 3.05 mg/mL, as determined by DPPH and ABTS assays. The antibacterial efficacy of these extracts was also tested against Streptococcus pyogenes, Streptococcus mutans, Staphylococcus aureus, and Klebsiella pneumoniae. The diameters of the inhibition zones ranged from 14.7 to 17.3 mm using the agar well diffusion method, with MIC and MBC values ranging from 7.81 to 250 mg/mL. Anti-biofilm formation, antibacterial adhesion, and antibacterial invasion assays further demonstrated that these medicinal plant extracts can inhibit bacterial biofilm formation and prevent the adhesion and invasion of oral pathogenic bacteria on the human tongue squamous cell carcinoma-derived cell line (HSC-4 cells). The ethanolic extracts of C. nutans and A. ebracteatus were able to inhibit the gtfD and gbp genes, which facilitate biofilm formation and bacterial adherence to surfaces. These findings provide new insights into the antibacterial and antioxidant properties of plant extracts from the Acanthaceae family. These activities could enhance the clinical and pharmaceutical applications of plant extracts as an alternative therapy for bacterial infections and a dietary supplement.

N-acetylcysteine effects on extracellular polymeric substances of Xylella fastidiosa: A spatiotemporal investigation with implications for biofilm disruption

BackgroundThe matrix of extracellular polymeric substances (EPS) present in biofilms greatly amplifies the problem of bacterial infections, protecting bacteria against antimicrobial treatments and eventually leading to bacterial resistance. The need for alternative treatments that destroy the EPS matrix becomes evident. N-acetylcysteine (NAC) is one option that presents diverse effects against bacteria; however, the different mechanisms of action of NAC in biofilms have yet to be elucidated. ObjectivesIn this work, we performed microscopy studies at micro and nano scales to address the effects of NAC at single cell level and early-stage biofilms of the Xylella fastidiosa phytopathogen. MethodsWe show the physical effects of NAC on the adhesion surface and the different types of EPS, as well as the mechanical response of individual bacteria to NAC concentrations between 2 and 20 mg/mL. ResultsNAC modified the conditioning film on the substrate, broke down the soluble EPS, resulting in the release of adherent bacteria, decreased the volume of loosely bound EPS, and disrupted the biofilm matrix. Tightly bound EPS suffered structural alterations despite no solid evidence of its removal. In addition, bacterial force measurements upon NAC action performed with InP nanowire arrays showed an enhanced momentum transfer to the nanowires due to increased cell mobility resulting from EPS removal. ConclusionsOur results clearly show that conditioning film and soluble EPS play a key role in cell adhesion control and that NAC alters EPS structure, providing solid evidence that NAC actuates mainly on EPS removal, both at single cell and biofilm levels.

Chitin-Assisted Synthesis of CuS Composite Sponge for Bacterial Capture and Near-Infrared-Promoted Healing of Infected Diabetic Wounds.

Diabetic wounds are prone to recurrent infections, often leading to delayed healing. To address this challenge, we developed a chitin-copper sulfide (CuS@CH) composite sponge, which combines bacterial trapping with near-infrared (NIR) activated phototherapy for treating infected diabetic wounds. CuS nanoparticles were synthesized and incorporated in situ within the sponge using a chitin assisted biomineralization strategy. The positively charged chitin surface effectively adhered bacteria, while NIR irradiation of CuS generated reactive oxygen species (ROS) heat and Cu2+ to rapidly damage the trapped bacteria. This synergistic effect resulted in an exceptional antibacterial performance against E. coli (∼99.9%) and S. aureus (∼99.3%). The bactericidal mechanism involved NIR-induced glutathione oxidation, membrane lipid peroxidation, and increased membrane permeability. In diabetic mouse models, the CuS@CH sponge accelerated the wound healing of S. aureus infected wounds by facilitating collagen deposition and reducing inflammation. Furthermore, the sponge demonstrated good biocompatibility. This dual-functional platform integrating bacterial capture and NIR-triggered phototherapy shows promise as an antibacterial wound dressing to promote healing of infected diabetic wound.

First biological response on polyAcrylic Acid UV-grafted PDMS surface: Towards medical application

Several biomedical devices and implants are made with Poly(dimethyl siloxane) (PDMS) or include it. Its biocompatibility, transparency, thermal stability and low cost are some of the properties that made PDMS suitable for biomedical applications. However, some properties such as its hydrophobic character can lead to complications owing to bacteria adhesion for instance. For breast implant, which is implanted for a long-time, an enhanced surface integration can be needed to avoid bacterial biofilm formation and degeneration. In microfluidic systems or for catheters, the biofouling phenomena must be avoided. Surface covalent functionalization with bioactive polymers is an attractive strategy to fight these issues. This study deals with poly(acrylic) acid UV-grafting on PDMS surfaces. After chemical and mechanical characterizations, this work has focused on biological response occurring on the functionalized material. The results show slowdown fibroblasts proliferation. Investigation around protein (BSA, fibronectin, fibrinogen) adhesion, made possible to approach surface mechanism understanding. FTIR and water contact angle analysis highlighted the different protein affinity towards surface. These results provide clues about protein orientation on the surface, in order to adapt the surface functionalization to target a specific application.

A flow cytometry method for quantitative measurement and molecular investigation of the adhesion of bacteria to yeast cells

The study of microorganism interactions is important for understanding the organization and functioning of microbial consortia. Additionally, the interaction between yeast and bacteria is of interest in the field of health and nutrition area for the development of probiotics. To investigate these microbial interactions at the cellular and molecular levels, a simple, reliable, and quantitative method is proposed. We demonstrated that flow cytometry enables the measurement of interactions at a single-cell level by detecting and counting yeast cells with bound fluorescent bacteria. Imaging flow cytometry revealed that the number of bacteria attached to yeast followed a Gaussian distribution whose maximum reached 14 bacterial cells using a clinical Escherichia coli strain E22 and the laboratory yeast strain BY4741. We found that the dynamics of adhesion resemble a Langmuir adsorption model, albeit it is a rapid and almost irreversible process. This adhesion is dependent on the mannose-specific type 1 fimbriae, as E. coli mutants lacking these appendages no longer adhere to yeast. However, this type 1 fimbriae-dependent adhesion could involve additional yeast cell wall factors, since the interaction between bacteria and yeast mutants with altered mannan content remained comparable to that of wild-type yeast. In summary, flow cytometry is an appropriate method for studying bacteria-yeast adhesion, as well as for the high-throughput screening of candidate molecules likely to promote or counteract this interaction.