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Biofilm-related ocular infections can lead to vision loss and are difficult to treat with antibiotics due to challenges with application and increasing microbial resistance. In turn, the design and testing of new synthetic drugs is a time- and cost-consuming process. Therefore, in this work, for the first time, we assessed the in vitro efficacy of the plant-based abietic acid molecule, both alone and when introduced to a polymeric cellulose carrier, against biofilms formed by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans in standard laboratory settings as well as in a self-designed setting using the topologically challenging surface of the artificial eye. These analyses were performed using the standard microdilution method, the biofilm-oriented antiseptic test (BOAT), a modified disk-diffusion method, and eyeball models. Additionally, we assessed the cytotoxicity of abietic acid against eukaryotic cell lines and its anti-staphylococcal efficacy in an in vivo model using Galleria mellonella larvae. We found that abietic acid was more effective against Staphylococcus than Pseudomonas (from two to four times, depending on the test applied) and that it was generally more effective against the tested bacteria (up to four times) than against the fungus C. albicans at concentrations non-cytotoxic to the eukaryotic cell lines and to G. mellonella (256 and 512 µg/mL, respectively). In the in vivo infection model, abietic acid effectively prevented the spread of staphylococcus throughout the larvae organisms, decreasing their lethality by up to 50%. These initial results obtained indicate promising features of abietic acid, which may potentially be applied to treat ocular infections caused by pathogenic biofilms, with higher efficiency manifested against bacterial than fungal biofilms.

Abstract The application of various magnetic fields for boosting the efficacy of different antimicrobial molecules or in the character of a self-reliant antimicrobial agent is considered a promising approach to eradicating bacterial biofilm-related infections. The purpose of this study was to analyze the phenomenon of increased activity of octenidine dihydrochloride-based antiseptic (OCT) against Staphylococcus aureus and Pseudomonas aeruginosa biofilms in the presence of the rotating magnetic field (RMF) of two frequencies, 5 and 50 Hz, in the in vitro model consisting of stacked agar discs, placed in increasing distance from the source of the antiseptic solution. The biofilm-forming cells' viability and morphology as well as biofilm matrix structure and composition were analyzed. Also, octenidine dihydrochloride permeability through biofilm and porous agar obstacles was determined for the RMF-exposed versus unexposed settings. The exposure to RMF or OCT apart did not lead to biofilm destruction, contrary to the setting in which these two agents were used together. The performed analyses revealed the effect of RMF not only on biofilms (weakening of cell wall/membranes, disturbed morphology of cells, altered biofilm matrix porosity, and composition) but also on its milieu (altered penetrability of octenidine dihydrochloride through biofilm/agar obstacles). Our results suggest that the combination of RMF and OCT can be particularly promising in eradicating biofilms located in such areas as wound pockets, where physical obstacles limit antiseptic activity.

The increasing bacterial drug resistance and the associated challenges in the treatment of infections warrant the search for alternative therapeutic methods. Hope is placed in antimicrobial peptides, which have a broad spectrum of action and are effective against strains which are resistant to conventional antibiotics. Antimicrobial peptides are also tested for their efficacy in the treatment of infections associated with the formation of biofilm. The aim of the present study was to examine the effect of Camel peptide on S. epidermidis and S. haemolyticus adhesion to and formation of biofilm on steel cortical bone screws and also on the process of reducing mature biofilm in orthopedic implants. The tests were performed on steel implants for osteosynthesis. The MIC value and MBEC values of the peptide were determined using the microdilution method in microtiter plates. The effect of the peptide on adhesion and biofilm formation, as well as on the activity on the preformed biofilm, was evaluated using quantitative methods and confocal microscopy. The presented research results indicate that the peptide exhibits very good antimicrobial properties against the analyzed strains. Concentrations above MIC reduced biofilm in the range of 90–99%.

In this paper we present the possibilities of using a femtosecond fiber laser (τ = 260 fs, λ = 1030 nm) to modify a PLLA surface. Three types of developed structures S1, S2, S3 differed significantly in topography and surface morphology. Thanks to the use of pulse overlapping in both scanning directions, the structures S1 and S2 were highly homogeneous, while in structure S3 the parallel grooves were obtained by pulse overlapping in one direction. The modified surfaces were examined for morphology (scanning electron microscopy), roughness (optical profilometry), wettability (measurement of the contact angle) and changes in the chemical structure of the polymer (infrared spectroscopy, gel permeation chromatography). The possibility of modifying these surface properties in a broad range was also discussed. The conducted cellular studies showed that there was no cytotoxicity with regard to all samples and that the pace of fibroblast colonization of samples S1 was higher compared to that of the control during the first 24 h of experiment. Moreover, the ability of S. aureus (pathogenic bacteria) and of a biotechnologically useful fungus Saccharomyces boulardii to colonize the manufactured structures was also assessed. In this case, no significant differences in the number of biofilm-forming cells were observed between the analyzed samples and the control setting. Obtained results indicate that applicability of such surface modifications should be further investigated, especially for the development of bone implants and these biotechnological processes in which microbes are used to produce the desired yield of product.

The processing of new generation titanium alloys using the Laser-based Powder Bed Fusion (L-PBF) technology is an innovation in the world of science. The selection of parameters as well as the determination of the process window enabling the processing of powder based on the chemical composition of Ti-13Nb-13Zr is the main element of this research. The wide spectrum of powder tests, such as batch powder analysis in the field of microscopic examination, powder fraction and distribution, dynamic flow tests, and examination of the chemical composition of powders, have delivered comprehensive results. Followed by the development of key set parameters affecting the efficiency of the L-PBF process and thus providing low porosity samples. As part of this study, an analysis of the influence of the thermal additive manufacturing process on the microstructural changes of the material was carried out using the methods of scanning electron microscopy and X-ray diffraction, as well as the analysis of X-ray fluorescence. The research confirmed the possibility of obtaining a high relative density of the material produced with the use of additive technologies at the level of over 99% while maintaining a stable chemical composition of the material meeting the ASTM F1713 conditions.

The disruption of the balance in the composition of oral and throat microbiota due to overgrowth of infectious biofilms may lead not only to severe infections, but also to serious, systemic complications resulting in a permanent loss of health or even in the death of the patient. Therefore, a number of hygienic measures are applied to counter-act such a threat, including the provision of locally active antiseptic agents. In this work, the antimicrobial activity of a well-established drug, octenidine dihydrochloride, in a new formulation of lozenges toward the biofilms formed in vitro by Staphylococcus aureus, Streptococcus pyogenes, Candida albicans, Pseudomonas aeruginosa and Aggregatibacter actinomycetemcomitans was assessed. The metabolic activity and quantitative culturing were performed and also scanning electron and confocal microscopies with regard to these biofilms exposed to the activity of octenidine in lozenges vs. a comparator (octenidine dihydrochloride dissolved in liquid). Biofilms were cultured in different experimental settings, including one in which hydroxyapatite served as the biofilm growth surface and using artificial saliva as the biofilm milieu. The obtained results indicated that the tested formulation of octenidine-containing lozenges displayed a high efficacy towards the analyzed biofilms, regardless of the in vitro setting applied. The current work is of a strictly in vitro nature, and the analyses were performed on reference microbial strains and not on the spectrum of clinical strains. Nevertheless, considering the fact that the time of exposition of biofilm to the octenidine released from the lozenge is significantly longer than the contact time of an antiseptic dissolved in liquid also tested in this study, it can be assumed that stable carriers of octenidine may find its broad application in the maintenance of oral and throat hygiene and the eradication of biofilm-based infections.

Hernia repairs are the most common abdominal wall elective procedures performed by general surgeons. Hernia-related postoperative infective complications occur with 10% frequency. To counteract the risk of infection emergence, the development of effective, biocompatible and antimicrobial mesh adjuvants is required. Therefore, the aim of our in vitro investigation was to evaluate the suitability of bacterial cellulose (BC) polymer coupled with gentamicin (GM) antibiotic as an absorbent layer of surgical mesh. Our research included the assessment of GM-BC-modified meshes’ cytotoxicity against fibroblasts ATCC CCL-1 and a 60-day duration cell colonisation measurement. The obtained results showed no cytotoxic effect of modified meshes. The quantified fibroblast cells levels resembled a bimodal distribution depending on the time of culturing and the type of mesh applied. The measured GM minimal inhibitory concentration was 0.47 µg/mL. Results obtained in the modified disc-diffusion method showed that GM-BC-modified meshes inhibited bacterial growth more effectively than non-coated meshes. The results of our study indicate that BC-modified hernia meshes, fortified with appropriate antimicrobial, may be applied as effective implants in hernia surgery, preventing risk of infection occurrence and providing a high level of biocompatibility with regard to fibroblast cells.

Methicillin-resistant strains of Staphylococcus aureus (MRSA) have developed resistance to most β-lactam antibiotics and have become a global health issue. In this work, we analyzed the impact of a rotating magnetic field (RMF) of well-defined and strictly controlled characteristics coupled with β-lactam antibiotics against a total of 28 methicillin-resistant and sensitive S. aureus strains. The results indicate that the application of RMF combined with β-lactam antibiotics correlated with favorable changes in growth inhibition zones or in minimal inhibitory concentrations of the antibiotics compared to controls unexposed to RMF. Fluorescence microscopy indicated a drop in the relative number of cells with intact cell walls after exposure to RMF. These findings were additionally supported by the use of SEM and TEM microscopy, which revealed morphological alterations of RMF-exposed cells manifested by change of shape, drop in cell wall density and cytoplasm condensation. The obtained results indicate that the originally limited impact of β-lactam antibiotics in MRSA is boosted by the disturbances caused by RMF in the bacterial cell walls. Taking into account the high clinical need for new therapeutic options, effective against MRSA, the data presented in this study have high developmental potential and could serve as a basis for new treatment options for MRSA infections.

In this research we subjected samples of poly(L-lactide) (PLLA) extruded film to ultraviolet (193 nm ArF excimer laser) radiation below the ablation threshold. The modified film was immersed in Simulated Body Fluid (SBF) at 37 °C for 1 day or 7 days to obtain a layer of apatite ceramic (CaP) coating on the modified PLLA surface. The samples were characterized by means of optical profilometry, which indicated an increase in average roughness (Ra) from 25 nm for the unmodified PLLA to over 580 nm for irradiated PLLA incubated in SBF for 1 day. At the same time, the water contact angle decreased from 78° for neat PLLA to 35° for irradiated PLLA incubated in SBF, which suggests its higher hydrophilicity. The obtained materials were investigated by means of cell response fibroblasts (3T3) and macrophage-like cells (RAW 264.7). Properties of the obtained composites were compared to the unmodified PLLA film as well as to the UV-laser irradiated PLLA. The activation of the PLLA surface by laser irradiation led to a distinct increase in cytotoxicity, while the treatment with SBF and the deposition of apatite ceramic had only a limited preventive effect on this harmful impact and depended on the cell type. Fibroblasts were found to have good tolerance for the irradiated and ceramic-covered PLLA, but macrophages seem to interact with the substrate leading to the release of cytotoxic products.

Biofilms are surface-attached, structured microbial communities displaying higher tolerance to antimicrobial agents in comparison to planktonic cells. An estimated 80% of all infections are thought to be biofilm-related. The drying pipeline of new antibiotics efficient against biofilm-forming pathogens urges the search for alternative routes of treatment. Essential Oils (EOs), extracted from medicinally important plants, are a reservoir of bioactive compounds that may serve as a foothold in investigating novel antibiofilm compounds. The aim of this study was to compare antimicrobial activity of liquid and volatile fractions of tested EOs against biofilm-forming pathogens using different techniques. In this research, we tested five EOs, extracted from Syzygium aromaticum L., Boswelia serrata Roxb., Juniperus virginiana L., Pelargonium graveolens L. and Melaleuca alternifolia Cheel., against planktonic and biofilm forms of five selected reference strains, namely Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. To obtain cohesive results, we applied four various methodological approaches: to assess the activity of the liquid fraction of EOs, disc diffusion and the microdilution method were applied; to test EOs’ volatile fraction, the AntiBioVol assay and modified Antibiofilm Dressing Activity Measurement (A.D.A.M.) were used. The molecular composition and dynamics of antimicrobial substances released from specific EOs was measured using Gas Chromatography–Mass Spectrometry (GC-MS). The antimicrobial potency of EO’s volatile fraction against biofilm formed by tested strains differed from that of the liquid fraction and was related to the molecular weight of volatile compounds. The liquid fraction of CW-EO and volatile fraction of F-EO acted in the strongest manner against biofilm of C. albicans. The addition of 0.5% Tween 20 to liquid phase, enhanced activity of G-EO against E. coli and K. pneumoniae biofilm. EO activity depended on the microbial species it was applied against and the chosen assessment methodology. While all tested EOs have shown a certain level of antimicrobial and antibiofilm effect, our results indicate that the choice of EO to be applied against a specific biofilm-forming pathogen requires careful consideration with regard to the above-listed aspects. Nevertheless, the results presented in this research contribute to the growing body of evidence indicating the beneficial effects of EOs, which may be applied to fight biofilm-forming pathogens.

In this research, we prepared foam scaffolds based on poly(l-lactide) (PLLA) and apatite whiskers (HAP) using thermally induced phase separation technique supported by the salt leaching process (TIPS-SL). Using sodium chloride having a size of (a) 150–315 μm, (b) 315–400 μm, and (c) 500–600 μm, three types of foams with different pore sizes have been obtained. Internal structure of the obtained materials has been investigated using SEM as well as μCT. The materials have been studied by means of porosity, density, and compression tests. As the most promising, the composite prepared with salt size of 500–600 μm was prepared also with the l-lysine modified apatite. The osteoblast hFOB 1.19 cell response for the scaffolds was also investigated by means of cell viability, proliferation, adhesion/penetration, and biomineralization. Direct contact cytotoxicity assay showed the cytocompatibility of the scaffolds. All types of foam scaffolds containing HAP whiskers, regardless the pore size or l-lysine modification induced significant stimulatory effect on the cal-cium deposits formation in osteoblasts. The PLLA/HAP scaffolds modified with l-lysine stimulated hFOB 1.19 osteoblasts proliferation. Compared to the scaffolds with smaller pores (150–315 µm and 315–400 µm), the PLLA/HAP foams with large pores (500–600 µm) promoted more effective ad-hesion of osteoblasts to the surface of the biomaterial.

The purpose of this work is to obtain comprehensive reference data of the Ti-13Nb-13Zr alloy base material: its microstructure, mechanical, and physicochemical properties. In order to obtain extensive information on the tested materials, a number of examination methods were used, including SEM, XRD, and XPS to determine the phases occurring in the material, while mechanical properties were verified with static tensile, compression, and bending tests. Moreover, the alloy’s corrosion resistance in Ringer’s solution and the cytotoxicity were investigated using the MTT test. Studies have shown that this alloy has the structure α’, α, and β phases, indicating that parts of the β phase transformed to α’, which was confirmed by mechanical properties and the shape of fractures. Due to the good mechanical properties (E = 84.1 GPa), high corrosion resistance, as well as the lack of cytotoxicity on MC3T3 and NHDF cells, this alloy meets the requirements for medical implant materials. Ti-13Nb-13Zr alloy can be successfully used in implants, including bone tissue engineering products and dental applications.

Electron beam melting (EBM) is an additive manufacturing technique, which allows forming customized implants that perfectly fit the loss of the anatomical structure of bone. Implantation efficiency depends not only on the implant’s functional or mechanical properties but also on its surface properties, which are of great importance with regard to such biological processes as bone regeneration or microbial contamination. This work presents the impact of surface modifications (mechanical polishing, sandblasting, and acid-polishing) of EBM-produced Ti6Al4V ELI implants on essential biological parameters. These include wettability, cytotoxicity toward fibroblast and osteoblast cell line, and ability to form biofilm by Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans. Obtained results indicated that all prepared surfaces exhibited hydrophilic character and the highest changes of wettability were obtained by chemical modification. All implants displayed no cytotoxicity against osteoblast and fibroblast cell lines regardless of the modification type. In turn, the quantitative microbiological tests and visualization of microbial biofilm by means of electron microscopy showed that type of implant’s modification correlated with the species-specific ability of microbes to form biofilm on it. Thus, the results of the presented study confirm the relationship between such technological aspects as surface modification and biological properties. The provided data are useful with regard to applications of the EBM technology and present a significant step towards personalized, customized implantology practice.