Direct Contact Assay Research Articles

Cytotoxicity and bactericidal activity of alginate/polyethylene glycol films with zinc oxide or silicon oxide nanoparticles for food packaging

In the food industry, packaging is used to protect food. However, traditional packaging has serious environmental impacts. Thus, studies have investigated packaging composed of biodegradable polymers. Accordingly, the present study aimed to obtain and characterize polymeric films of sodium alginate and polyethylene glycol crosslinked with tetraethoxysilane and doped with nano-zinc oxide or nano-silicon oxide and evaluate their cytotoxicity and bactericidal activity against coliforms and Escherichia coli. The nanoparticles and crosslinked materials were characterized using infrared and Raman spectroscopy. The cytotoxicity of the films was evaluated using a direct contact assay with a bovine fibroblast cell model. The bactericidal activity of the films was assessed by determining whether the contamination indicators were activated. The average sizes of the ZnO and SiO2 nanoparticles were 408.3 and 125.3 nm, respectively, and their zeta potentials were −31.6 and −31.1 mV, respectively. Cell growth inhibition zone tests indicated that 75 ALG/25 PEG/SiO2 was slightly toxic, while 25 ALG/75 PEG/ZnO exhibited mild toxicity. Cell area analysis showed that the addition of SiO2 decreased the cytotoxicity of the obtained films. Finally, it was possible to observe the inactivation of both contamination bioindicators. Therefore, films containing nanoparticles could be promising biomaterials for packaging in the food industry.

Whole-cell biosensors for determination of bioavailable pollutants in soils and sediments: Theory and practice

The bioavailability of pollutants is a key factor affecting environmental risk. Whole-cell bioreporters are a demonstratedly effective tool for the investigation of pollutant bioavailability in water and soil/sediment. Unlike aqueous samples, transmittance of bioreporter optical signal is reduced in direct-contact assays with soil/sediment, which affects the accuracy of bioreporter-detected pollutant bioavailability. No studies have measured the magnitude and variability of soil/sediment effects on signal in direct-contact assays or how associated uncertainties influence results. In this study, we investigate the optical effects of soil/sediment particles in suspensions on bioreporter signal transmittance and quantify how variable these optical effects are from sample-to-sample. We find that neglecting bioreporter signal diminution by soil/sediment, as many studies do, can lead to order-of-magnitude errors in results, underestimating risk. Correction based on methods in ad hoc use (e.g. comparison to signal from non-inducible reporter or use of reference soil/sediment) are also problematic for some types of experiment, and could lead to errors in excess of 30%. Our findings have a sound basis in theory, and we provide recommendations concerning the most suitable type of approach to use for different experimental settings. Generally, if best accuracy is not needed to quantify bioavailability, for samples that have been ground, sieved, and are of reasonably uniform color, it may be possible to use a single or average correction factor, particularly for experiments performed at a single slurry concentration. For investigations studying bioavailability under varying solid-phase:water ratios (e.g., sorption/desorption), detailed compensation measurements are needed for independent variables, including each specific soil/sediment sample, slurry concentration, and in some cases bioreporter signal intensity. Our measurements and calculations indicate that best results are obtained when working in the region of ballistic photon transmittance. Findings herein will be useful in areas that require information on bioavailability, such as ecotoxicology and environmental risk assessment.

Evaluation of the Antimicrobial Effect of Mineral Trioxide Aggregate Mixed with Fluorohydroxyapatite against E. faecalis In Vitro

Enterococcus faecalis is the dominant microorganism in chronic apical periodontitis. It is more resistant to local antiseptic agents than other endodontic microorganisms. Currently, mineral trioxide aggregate (MTA) is considered as an ideal material in many endodontic procedures. Some studies have shown that MTA has good antibacterial activity against E. faecalis. However, some studies have investigated the effect of incorporating some materials into MTA on its antibacterial activity against E. faecalis. No study has evaluated the effect of incorporating fluorohydroxyapatite nanoparticles (nano-FHA) on the antimicrobial activity of MTA. Therefore, the present study evaluated the antimicrobial effect of MTA mixed with nano-FHA on E. faecalis in vitro. The study was carried out on 18 samples in three groups: pure MTA, MTA mixed with 10 wt% of nano-FHA, and MTA mixed with 15 wt% of nan-FHA. The effect of nano-FHA on the antibacterial activity of MTA on E. faecalis was evaluated by evaluating the growth inhibition zone around each sample. The antimicrobial effect of samples on inhibiting E. faecalis biofilm formation and inhibiting microbial growth of E. faecalis in the planktonic phase was evaluated by disk agar diffusion (DAD), biofilm inhibition assay (BIA), and direct contact assay (DCA) tests, respectively. All the above tests were analyzed after 24 and 72 hours. Factorial designs were used for statistical analyses. Tukey tests were used for two-by-two comparisons. All the statistical analyses were carried out with SPSS 26. DAD results showed no formation of the growth inhibition zone in all the samples after 24 and 72 hours. The microbial colony counts in the BIA and DCA tests in the groups modified with FHA nanoparticles were significantly lower than the pure MTA group (P < 0.05). The microbial colony counts increased in all the groups over time (P < 0.05). Incorporating nano-FHA into MTA improved the antimicrobial activity of MTA against E. faecalis compared to pure MTA. The highest antimicrobial activity was achieved after incorporating 15 wt% of nano-FHA into MTA at the 72-hour interval.

The myth of antibiotic spider silk

SummarySpider silk is frequently attributed antimicrobial properties. This notion is based on studies reporting antimicrobial activity (AMA) of spider silk; however, close inspection of these studies reveals that the evidence is conflicting, and at best anecdotal. We performed a systematic study of antimicrobial properties of different silk types from seven species across the spider phylogeny. We found no evidence of AMA of silk in direct contact and disc diffusion assays against Gram-negative Escherichia coli and Pseudomonas putida, and the Gram-positive Bacillus subtilis. Furthermore, staining experiments and fluorescence microscopy showed the presence of live bacteria on silk surfaces indicating no antimicrobial effect on direct contact. A critical evaluation of the literature reveals that published tests of AMA are scarce and that all the studies claiming positive results are compromised by methodological shortcomings. Our analysis demonstrates that the common notion that spider silk is antimicrobial is not supported by empirical data.

Accelerated biodegradation of iron-based implants via tantalum-implanted surface nanostructures

In recent years, pure iron (Fe) has attracted significant attention as a promising biodegradable orthopedic implant material due to its excellent mechanical and biological properties. However, in physiological conditions, Fe has an extremely slow degradation rate with localized and irregular degradation, which is problematic for practical applications. In this study, we developed a novel combination of a nanostructured surface topography and galvanic reaction to achieve uniform and accelerated degradation of an Fe implant. The target-ion induced plasma sputtering (TIPS) technique was applied on the Fe implant to introduce biologically compatible and electrochemically noble tantalum (Ta) onto its surface and develop surface nano-galvanic couples. Electrochemical tests revealed that the uniformly distributed nano-galvanic corrosion cells of the TIPS-treated sample (nano Ta–Fe) led to relatively uniform and accelerated surface degradation compared to that of bare Fe. Furthermore, the mechanical properties of nano Ta–Fe remained almost constant during a long-term in vitro immersion test (~40 weeks). Biocompatibility was also assessed on surfaces of bare Fe and nano Ta–Fe using in vitro osteoblast responses through direct and indirect contact assays and an in vivo rabbit femur medullary cavity implantation model. The results revealed that nano Ta–Fe not only enhanced cell adhesion and spreading on its surface, but also exhibited no signs of cellular or tissue toxicity. These results demonstrate the immense potential of Ta-implanted surface nanostructures as an effective solution for the practical application of Fe-based orthopedic implants, ensuring long-term biosafety and clinical efficacy.

Antimicrobial activity of AH Plus and Ceaseal sealers with and without the addition of silver nanoparticles against Enterococcus faecalis.

Aim: The aim of the present study was to investigate the antimicrobial effect of AH Plus and Ceraseal sealers with and without the incorporation of silver nanoparticles against Enterococcus faecalis using the direct contact assay. Methodology: Silver nanoparticles were prepared by chemical reduction method. Direct Contact Test (DCT) was carried out by settling 50mg of freshly mixed sealer in a sterile flat bottom screw capped tube.Ten tubes were prepared for each sealer in duplicate as follow:Group(I): Ceraseal (Meta Biomed Co., Cheongju, Korea) , Group (II): Ceraseal& Nano silver gel, Group(III): AH Plus (Dentsply/Maillefer, Konstanz, Germany) and Group (IV): AH Plus & Nano silver gel. Antibacterialactivity was evaluated against Enterococcus faecalis ATCC 35550 at various time intervals 1 ,24 and 168 hours,. Results: DCT results revealed that AH Plus had higher antibacterial activity against Enterococcus faecalis at different time intervals than Ceraseal sealer unaffected by (AgNPs)incorporation. But the incorporation of(AgNPs)significantly increased their antibacterial effect. All sealers had lost much of their antibacterial effects at 7 days interval. Conclusion: AH Plus sealer had higher antibacterial effect than Ceraseal sealer. Incorporation of (AgNPs) silver nanoparticles can improve the antibacterial activity of AH Plus and Ceraseal sealer.

Antimicrobial properties of dental cements modified with zein-coated magnesium oxide nanoparticles

The aim of this study was to test the antimicrobial properties of dental cements modified with magnesium oxide (MgO) nanoparticles. Zein-modified MgO nanoparticles (zMgO) in concentrations (0.0, 0.3, 0.5, and 1.0%) were mixed with dental cements (Fuji II, Rely X Temp E, Ionoglass Cem, Es Temp NE, and System P link). Eight discs were fabricated from each zMgO-cement pair for a total of 32 specimens for each cement. Characterization of the dental cements incorporating zMgO was done by X-ray Diffraction (XRD) and Field Emission Scanning Electron Microscopy (FESEM). The antimicrobial properties of the mixtures were tested using direct contact and agar diffusion assays against Streptococcus mutans, Staphylococcus aureus, Enterococcus faecalis, and Candida albicans. Data was analyzed using two-way analysis of variance and LSD post hoc test at 0.05 significance level. XRD spectra showed sharp peaks of zMgO indicating its high crystalline nature, while the amorphous dental cements with zMgO had broad peaks. FESEM analysis showed a uniform distribution of the zMgO nanoparticles in the cement. There were significant inhibition zone values associated with all concentrations of zMgO-cement mixtures tested compared to controls (p < 0.001) with a dose-response recorded only with Fuji II. Optical density values were significantly lower in zMgO groups compared to controls for all microorganisms. The effect was most prominent with Rely X against C. albicans and S. aureus. Dental cements containing zMgO showed significant antimicrobial properties that were dependent on the specific initial cement substrate.

The effect of surface modification of TiMg composite on the in-vitro degradation response, cell survival, adhesion, and proliferation

This study is aimed to evaluate the influence of mechanical surface treatment on the degradation response, cell survival, adhesion, and proliferation of a TiMg composite material. Two sets of the TiMg samples with different surface characteristics were studied: i) as-machined samples (TiMg-T) and ii) samples with a mechanically modified surface (TiMg-P). Surface roughness was determined using a confocal microscope. Degradation rates (DR) were evaluated in artificial Plasma, HBSS, and NaCl 0.9%. The cell viability was evaluated using an MTT assay. The initial cell adhesion and spreading were investigated using the direct contact assay. An xCELLigence system was employed to provide real-time cell proliferation. The focal adhesion and cell morphological changes were also examined. The DR of TiMg-P decreased by ⁓5 times compared with that of TiMg-T. Surface of the TiMg-P specimens after 72 h exposure to either HBSS or Plasma was passivated by a layer enriched with bioactive Ca/P species. The cell viability of L929 and Saos-2 after 72 h incubation for TiMg-P was 94.6% and 94.8% compared with 73.8% and 74.3% obtained for TiMg-T, respectively. The direct contact assay showed that the initial adhesion and spreading of the L929 cells incubated with TiMg-P was more pronounced compared with that of TiMg-T. The proliferation rate of Saos-2 cells incubated with TiMg-P was higher when compared with that of TiMg-T, and was almost comparable to that of the DMEM-blank between the 24 and 72 h interval. TiMg-P had a pronounced difference in the number and area of Focal Adhesions (FA) compared with that of TiMg-T. The morphology of cells incubated with TiMg-P was not altered. The results confirmed that the smooth and less strained surface of the TiMg-P samples effectively improved the in-vitro degradation response, cell survival, adhesion, and proliferation.

Fabrication of Ca and P containing niobium oxide ceramic coatings on niobium by PEO coupled EPD process

Abstract Surface modification of niobium (Nb) to develop Ca and P containing niobium oxide (Nb2O5) ceramic coatings by plasma electrolytic oxidation (PEO) coupled with electrophoretic deposition (EPD) in a single step process is reported in the present study. Four coatings were obtained in silicate and phosphate electrolytes with and without the addition of hydroxyapatite (HA) nanoparticles. The phase composition, surface and cross-sectional morphology of the coatings were analysed. The corrosion resistance of the coated samples and niobium substrate was analysed using potentiodynamic polarisation (PDP) method in simulated body fluid (SBF) solution. The cell-viability and the live and dead cell morphology on the substrate and the coated samples were evaluated using L-929 cells. The adherence and growth of the L-929 cells were estimated by the direct contact assay. The surface modification by the PEO process enhanced the corrosion resistance of niobium in a fluid body environment. Among the coated samples, the PEO-EPD samples obtained in silicate electrolyte exhibited better corrosion resistance (6.02 × 10−7 mA/cm2), optimal surface properties, lower contact angle (47°), higher surface energy (50 mJ/m2), good adhesion strength (Lc = 18.1 N) and better cell viability.

Chemical constituents of essential oils from Thymus vulgaris and Cymbopogon citratus and their insecticidal potential against the tomato borer, Tuta absoluta (Lepidoptera: Gelechiidae)

This study aimed to determine the chemical composition and insecticidal activity of essential oils from Thymus vulgaris (thyme) and Cymbopogon citratus (lemongrass) against the devastating pest, Tuta absoluta. Essential oils were extracted by hydrodistillation and chemically analyzed by Gas chromatography coupled to mass spectrometry. Their knockdown (KD50) and larvicidal (LD50) activities were assessed through direct contact and fumigation toxicity assays. The major components of thyme oil were Thymol (21.53%), α-Pinene (Dextro) (17.43%) and o-Cymene (15.37%), while, Neral (34.48%), Geranial (34.37%) and β-Myrcene (12.84%) were prominent in lemongrass oil. Statistical analysis indicated that both oils exhibited similar knockdown and insecticidal efficiencies through direct contact and fumigation routes. The resultant biological parameters for lemongrass and thyme oils were KD50 values of 0.193 µL/mL and 0.592 µL/mL and LD50 values of 0.328 µL/mL and 0.608 µL/mL respectively for contact toxicity and KD50 of 1.290 µL/mL and 2.565 µL/mL and LD50 of 1.479 µL/mL and 3.046 µL/mL for lemongrass and thyme oils respectively for fumigant toxicity. The reference insecticide, Lynx (λ-cyhalothrin 15 g/L + acetamiprid 20 g/L) elicited no mortality through both routes after 4 hours of exposure. Both essential oils prolonged life cycle of insect pest through both routes as compared to the reference insecticide. However, greenhouse and open field trials are required to validate this approach prior to its implementation for T. absoluta control in tomato farms. To our knowledge, this study reports for the first time the insecticidal potential of lemongrass essential oil against T. absoluta.

Physical and biological characterizations of TiNbSn/(Mg) system produced by powder metallurgy for use as prostheses material

Titanium scaffolds with non-toxic β stabilizing elements (Nb and Sn), Ti–34Nb–6Sn (TNS), and with magnesium as spacer (TNS/M), were processed by powder metallurgy, and sintered at 800 °C. The X-ray diffraction (XRD) pattern showed that materials are biphasic alloys, presenting 45 to 42% (wt %) in hcp (α-phase) and the rest is bcc (β-phase), and the presence of a slight peak relating to TiO2 in both materials. Pores of approximately 50 μm for TNS and 300 μm to TNS/M were observed in the micrographic analysis by scanning electron microscopy (SEM). The wettability was higher for TNS/M compared to TNS. The elastic modulus was higher for TNS compared to TNS/M. Stem cells derived from equine bone marrow (BMMSCs) were used for in vitro assays. The morphologic and adhesion evaluation after 72 h, carried out by direct contact assay with the materials showed that the BMMSCs were anchored and adhered to the porous scaffolds, in the way the cytoplasmic extension was observed. The cellular migration, using the “wound healing” method, was significant for the groups treated with conditioned medium with materials in 24 h. Osteogenic differentiation of BMMSCs, assessed by calcium deposition and staining with Alizarin Red, was greater in the conditioned medium with TNS/M in 10 days of culture. Since the biological effects was good and the elastic modulus decreased in the system with magnesium is a promising new content titanium alloy for biomedical application.

Fabrication of ceramic coatings on the biodegradable ZM21 magnesium alloy by PEO coupled EPD followed by laser texturing process

Abstract Zirconia (ZrO2) incorporated ceramic coatings were fabricated on biodegradable ZM21 Mg alloy by the PEO coupled with EPD process. Subsequently, the sample surface was modified by laser texturing to improve the corrosion resistance, roughness and cell proliferation and growth properties. The corrosion performance of the fabricated samples along with the substrate was studied by electrochemical measurements under simulated body fluid (SBF) environment. The cell direct contact assay was conducted for the substrate and fabricated samples using L -929 mouse fibroblast cells for 24 h. The phase contrast images of cell direct contact assay revealed that fabricated samples exhibited better contact and response with the fibroblast cells, compared to the substrate. The addition of nanoparticles in the PEO process, called PEO coupled EPD process, resulted in attaining a higher thickness and improved corrosion performance of the samples than the PEO coated samples. Among all the samples, laser surface textured PEO, and PEO-EPD coated samples unveiled enhanced corrosion resistance, cell growth, thereby enabling it as a suitable prototype for biodegradable implant applications.

The antifungal activity of vapour phase of odourless thymol derivate.

Thymol is a substance with a great therapeutic potential possessing antibacterial and antifungal activity, with a characteristic odour that remains long after application but is not pleasant at higher concentrations. In this study, attention has been focused on describing the chemical and biological properties of the simply prepared trimethylsilyl ether of thymol (kubicin). Interestingly, kubicin has similar volatility as thymol, undergoes hydrolysis in the water (moisture; forming thymol and trimethylsilanol) and can be used at 6,000 times higher concentration than thymol without any negative and irritating odour. Kubicin showed diverse fungistatic and fungicidal activities when tested by direct contact assay, or in vapour phase. The volatile vapour of kubicin was effective on all tested fungal strains. These results suggest that vapours of kubicin might provide an alternative way to fight against fungal contamination.

Antifungal and Phytotoxic Activities of Essential Oils: In Vitro Assays and Their Potential Use in Crop Protection

(1) Background: The use of natural products based on essential oils (EO) is nowadays arousing great interest as an alternative method to control plant pathogens and weeds. However, EO possess low bioavailability and are highly volatile, and their encapsulation in hydroxypropyl-ß-cyclodextrin (HP-β-CD) could be a means to enhance their stability and maintain their bioactivity. Thus, the current study aims at investigating, in the presence and the absence of HP-β-CD, the antifungal and phytotoxic activities of nine EO, distilled from plant species belonging to Alliaceae, Apiaceae, and Cupressaceae families, with considerations for their chemical composition. (2) Methods: EO antifungal activity was assessed by direct contact and volatility assays against Fusarium culmorum, a major phytopathogenic fungi, while phytotoxic effects were evaluated against lettuce (Lactuca sativa L.) and rye-grass (Lolium perenne L.), by seedling’s emergence and growth assays. (3) Results: These EO inhibit fungal growth in both direct contact and volatility assays, with half-maximal inhibitory concentrations (IC50) ranging from 0.01 to 4.2 g L−1, and from 0.08 up to 25.6 g L−1, respectively. Concerning phytotoxicity, these EO have shown great potential in inhibiting lettuce (IC50 ranging from 0.0008 up to 0.3 g L−1) and rye-grass (IC50 ranging from 0.01 to 0.8 g L−1) seedlings’ emergence and growth. However, the EO encapsulation in HP-β-CD has not shown a significant improvement in EO biological properties in our experimental conditions. (4) Conclusion: All tested EO present antifungal and phytotoxic activities, with diverse efficacy regarding their chemical composition, whilst no increase of their biological effects was observed with HP-β-CD.

Synthesis and property of polyurethane elastomer for biomedical applications based on nonaromatic isocyanates, polyesters, and ethylene glycol

Polyurethane (PU) elastomers were synthesized by the reaction of HDI or IPDI diisocyanates and poly(ε-caprolactone) (PCL or poly(ethylene adipate) (PA) diols and ethylene glycol as a polymer chain extender. IR, 1H, and 13C NMR spectroscopy and X-ray analysis were used for the structural analysis of the formed films. The molecular weight distribution was examined by GPC chromatography. Based on the measured contact angles, free surface energy parameters were calculated. The obtained results were analyzed for the possible use of these polyurethanes as biomaterials. The most promising in this respect was PU-3, which was synthesized from IPDI and PCL. This was due to its high molecular weight of approximately 90,000, the presence of a crystalline phase, and the relatively high hydrophobicity, with a SEP value below 25 mJ/m2. These films showed a good resistance to hydrolysis during incubation in Baxter physiological saline during 6 weeks. Both Gram-positive (Bacillus sp.) and Gram-negative (Pseudomonas sp.) types of bacterial strains were used to test the biodegradation property. Synthesized PUs are biodegradable and showed moderate or even mild cytotoxicity against human normal fibroblasts (BJ) and immortalized keratinocytes (HaCaT), estimated with direct contact assay. The most biocompatible was PU-3 film, which revealed rather mild reactivity against both cell lines, and the least was PU-2 film, synthesized from HDI and PA (severe toxicity for HaCaTs).

Cytotoxicity and inflammatory response of different types of provisional restorative materials

AimTo evaluate the cytotoxicity and inflammatory response of different types of provisional restorative materials to mice gingival fibroblasts. MethodsCytotoxicity of provisional material discs (thermal-polymerized acrylic resin; auto-polymerized acrylic resin; bisacrylic resin; nano-ceramic resin for CAD/CAM and prefabricated polymer block for CAD/CAM) to Mice (Balb/c) gingival cell were investigated under direct and indirect contact (extracts) at 24, 48 and 72 h, using the MTT and Alamar blue assays. Materials extracts (24 h) were applied to the cell culture (indirect contact) or cells were seeded on discs of provisional materials, and the cytotoxicity and production of IL-6, IL-1β and TNF-α after 24, 48 and 72 h were analyzed through MTT, Alamar Blue® and ELISA. Culture medium was used as control for indirect contact assay (extract) and the surfaces of the wells without discs of provisional materials were used as control for direct contact assay. Results were analysed statistically by ANOVA followed by the Bonferroni-Test correction. Statistically significant differences were considered if P was < .05. ResultsAuto-polymerized and bisacrylic resins (direct contact) reduced cell viability after 24, 48 and 72 h compared to control (P < .05). Indirect contact (extract) was not cytotoxic to cells at all periods compared to control (P > .05). Auto-polymerized and bisacrylic resins increased IL-6, IL-1β and TNF-α levels mainly at 24 h when compared to the other materials (P < .001). ConclusionAuto-polymerized and bisacrylic were more cytotoxic to mice gingival fibroblasts. CAD/CAM nano ceramic resin and prefabricated polymer blocks are more predictable materials to preserve the periodontal soft tissues.

Low-density polyethylene films incorporated with silver nanoparticles to promote antimicrobial efficiency in food packaging.

Technological innovations in packaging are intended to prevent microbiological contaminations for ensuring food safety and preservation. In this context, researchers have investigated the antimicrobial effect of low-density polyethylene films incorporated with the following concentrations of silver nanoparticles: 1.50, 3.75, 7.50, 15.00, 30.00, 60.00, and 75.00 µg/ml. The films were characterized using field emission gun scanning electron microscopy, Fourier transform infrared spectroscopy, X-ray diffraction, thermogravimetry, and differential scanning calorimetry. From the results of these techniques, it could be concluded that the silver nanoparticles incorporated in the low-density polyethylene films did not influence their physical, chemical, and thermal properties. The direct contact assays, shake-flask assays, and bacterial images obtained using scanning electron microscopy were used to analyze the antimicrobial activity of the films. In the microbial analyses, it was verified that the nanostructured films exhibited antimicrobial properties against all the microorganisms studied, although more notably for fungi and Gram-negative bacteria than the Gram-positive bacteria. Moreover, it was discovered that the packages, in which silver nanoparticles were incorporated, inhibited the growth and reproduction of bacterial cells during the early stages. These results suggest that the extruded low-density polyethylene films incorporated with silver nanoparticles may be an essential tool for improving food quality and safety.

Sterilization efficiency of pathogen-contaminated cottons in a laundry machine.

Pathogenic bacteria on abiotic surfaces such as fabrics, bedding, patient wears, and surgical tools are known to increase the risk of bacterial diseases in infants and the elderly. The desiccation tolerance of bacteria affects their viability in cotton. Thus, washing and drying machines are required to use conditions that ensure the sterilization of bacteria in cotton. The objective of this study is to determine the effects of various sterilization conditions of washing and drying machines on the survival of three pathogenic bacteria (Acinetobacter baumannii, Pseudomonas aeruginosa, and Staphylococcus aureus) commonly presented in contaminated cotton and two non-pathogenic bacteria (Bacillus subtilis and Escherichia coli) in cotton. High survival rates of A. baumannii and S. aureus in desiccated cotton were observed based on scanning electron microscope and replicate organism direct agar contact assay. The survival rates of A. baumannii and S. aureus exposed in desiccated cotton for 8 h were higher (14.4 and 5.0%, respectively) than those of other bacteria (< 0.5%). All tested bacteria were eradicated at low-temperature (< 40°C) washing with activated oxygen bleach (AOB). However, bacterial viability was shown in low temperature washing without AOB. High-temperature (> 60°C) washing was required to achieve 99.9% of the sterilization rate in washing without AOB. The sterilization rate was 93.2% using a drying machine at 60°C for 4 h. This level of sterilization was insufficient in terms of time and energy efficiency. High sterilization efficiency (> 99.9%) at 75°C for 3 h using a drying machine was confirmed. This study suggests standard conditions of drying machines to remove bacterial contamination in cotton by providing practical data.

Burst Release of Antibiotics Combined with Long-Term Release of Silver Targeting Implant-Associated Infections: Design, Characterization and in vitro Evaluation of Novel Implant Hybrid Surface.

Implant-associated infections represent a serious risk in human medicine and can lead to complications, revisions and in worst cases, amputations. To target these risks, the objective was to design a hybrid implant surface that allows a local burst release of antibiotics combined with long-term antimicrobial activity based on silver. The efficacy should be generated with simultaneous in vitro cytocompatibility. The investigations were performed on titanium K-wires and plates and gentamicin was selected as an illustrative antibiotic. A gentamicin depot (max 553 µg/cm2) was created on the surface using laser structuring. The antibiotic was released within 15 min in phosphate buffered saline (PBS) or agar medium. Metallic silver particles (4 µg/cm2) in a titanium dioxide layer were deposited using plasma vapor deposition (PVD). About 16% of the silver was released within 28 days in the agar medium. The local efficacy of the incorporated silver was demonstrated in a direct contact assay with a reduction of more than 99.99% (Escherichia coli). The local efficacy of the hybrid surface was confirmed in a zone of inhibition (ZOI) assay using Staphylococcus cohnii. The biocompatibility of the hybrid surface was proven using fibroblasts and osteoblasts as cell systems. The hybrid surface design seems to be promising as treatment of implant-associated infections, considering the achieved amount and release behavior of the active ingredients (gentamicin, silver). The generated in vitro results (efficacy, biocompatibility) proofed the concept. Further in vivo studies will be necessary translate the hybrid surface towards clinical applied research.

The effect of aluminum oxide on red blood cell integrity and hemoglobin structure at nanoscale

Herein, we explored the interaction of Al2O3 NPs with RBCs and Hb to determine the effect of Al2O3 NPs on hemolytic activity and Hb denaturation. The percentage of hemolysis of extracts and direct contact assays triggered by Al2O3 NPs was calculated by determining supernatant Hb concentration at 540 nm. Far-UV CD and Trp/ANS/acrylamide fluorescence spectroscopic methods were used to determine the structural changes of Hb upon interaction with Al2O3 NPs. Theoretical studies were carried out to display the residues involved in the binding site of Hb with Al2O3 nanocluster as well as the structural changes of Hb after interaction. The results showed that the percentage of hemolysis of extract and direct contact assays induced by Al2O3 NPs were 1.16 and 0.46, respectively. Fluorescence spectroscopy revealed that Al2O3 NPs alter the quaternary structure of the protein; however, CD spectroscopy indicated that the secondary structure of Hb remains almost unchanged. Theoretical study displayed that Al2O3 nanocluster interacts with different residues of protein, and Hb tends to be destabilized at the binding site with nanocluster. This study may be significant in exploring the toxicity profile of Al2O3 NPs for their in vivo implementations.