Increase of ibuprofen penetration through the skin by forming ion pairs with amino acid alkyl esters and exposure to the electromagnetic field.

A method of increasing the permeability of ibuprofen through the skin using a rotating magnetic field (RMF) is presented. This study evaluated whether 50 Hz RMF modifies ibuprofen's permeability through the skin. Ibuprofen and its structural modifications in the form of ibuprofenates of isopropyl esters of L-amino acids such as L-valine, L-phenylalanine, L-proline, and L-aspartic acid were used in the research. To this end, Franz cells with skin as membrane were exposed to 50 Hz RMF with 5% ibuprofen and its derivatives in an ethanol solution for 48 h. Following the exposures, the amount of penetrated compound was analysed. Regardless of the compound tested, a significant increase in drug transport through the skin was observed. The differences in the first 30 min of permeation are particularly noticeable. Furthermore, it was shown that using RMF increases the permeability of ibuprofen from 4 to 244 times compared to the test without the RMF. The greatest differences were observed for unmodified ibuprofen. However, it is noteworthy that the largest amounts of the active substance were obtained with selected modifications and exposure to RMF. The RMF may be an innovative and interesting technology that increases the penetration of anti-inflammatory and anti-ache drugs through the skin.

Anticancer properties of bacterial cellulose membrane containing ethanolic extract of Epilobium angustifolium L.

Epilobium angustifolium L. is a medicinal plant well known for its anti-inflammatory, antibacterial, antioxidant, and anticancer properties related to its high polyphenols content. In the present study, we evaluated the antiproliferative properties of ethanolic extract of E. angustifolium (EAE) against normal human fibroblasts (HDF) and selected cancer cell lines, including melanoma (A375), breast (MCF7), colon (HT-29), lung (A549) and liver (HepG2). Next, bacterial cellulose (BC) membranes were applied as a matrix for the controlled delivery of the plant extract (BC-EAE) and characterized by thermogravimetry (TG), infrared spectroscopy (FTIR), and scanning electron microscopy (SEM) images. In addition, EAE loading and kinetic release were defined. Finally, the anticancer activity of BC-EAE was evaluated against the HT-29 cell line, which presented the highest sensitivity to the tested plant extract (IC50 = 61.73 ± 6.42 µM). Our study confirmed the biocompatibility of empty BC and the dose and time-dependent cytotoxicity of the released EAE. The plant extract released from BC-2.5%EAE significantly reduced cell viability to 18.16% and 6.15% of the control values and increased number apoptotic/dead cells up to 37.53% and 66.90% after 48 and 72 h of treatment, respectively. In conclusion, our study has shown that BC membranes could be used as a carrier for the delivery of higher doses of anticancer compounds released in a sustained manner in the target tissue.

Exposure to a rotating magnetic field as a method of increasing the skin permeability of active pharmaceutical ingredients.

The paper presents a method of increasing the permeability of various active substances through the skin by means of a rotating magnetic field. The study used 50 Hz RMF and various active pharmaceutical ingredients (APIs) such as caffeine, ibuprofen, naproxen, ketoprofen, and paracetamol. Various concentrations of active substance solutions in ethanol were used in the research, corresponding to those in commercial preparations. Each experiment was conducted for 24 h. It was shown that, regardless of the active compound used, an increase in drug transport through the skin was observed with RMF exposure. Furthermore, the release profiles depended on the active substance used. Exposure to a rotating magnetic field has been shown to effectively increase the permeability of an active substance through the skin.

The use of the electromagnetic field in microbial process bioengineering.

An electromagnetic field (EMF) has been shown to have various stimulatory or inhibitory effects on microorganisms. Over the years, growing interest in this topic led to numerous discoveries suggesting the potential applicability of EMF in biotechnological processes. Among these observations are stimulative effects of this physical influence resulting in intensified biomass production, modification of metabolic activity, or pigments secretion. In this review, we present the current state of the art and underline the main findings of the application of EMF in bioprocessing and their practical meaning in process engineering using examples selected from studies on bacteria, archaea, microscopic fungi and yeasts, viruses, and microalgae. All biological data are presented concerning the classification of EMF. Furthermore, we aimed to highlight missing parts of contemporary knowledge and indicate weak spots in the approaches found in the literature.

PhageScore-based analysis of Acinetobacter baumannii infecting phages antibiotic interaction in liquid medium.

The growing interest in bacteriophages and antibiotics' combined use poses new challenges regarding this phenomenon's accurate description. This study aimed to apply the PhageScore methodology to assess the phage-antibiotic combination activity in liquid bacterial culture. For this purpose, previously described Acinetobacter infecting phages vB_AbaP_AGC01, Aba-1, and Aba-4 and antibiotics (gentamicin, ciprofloxacin, meropenem, norfloxacin, and fosfomycin) were used to obtain a lysis curve of bacteriophages under antibiotic pressure. The experimental data were analyzed using the Fractional Inhibitory Concentration Index (FICI) and PhageScore methodology. The results obtained by this method clearly show differences between phage lytic activity after antibiotic addition. Thus, we present the potential use of the PhageScore method as a tool for characterizing the phage antibiotic synergy in liquid culture. Further, the optimization of the PhageScore for this purpose can help compare antibiotics and their outcome on bacteriophage lytic activity.

Rotating Magnetic Field-Assisted Reactor Enhances Mechanisms of Phage Adsorption on Bacterial Cell Surface.

Growing interest in bacteriophage research and use, especially as an alternative treatment option for multidrug-resistant bacterial infection, requires rapid development of production methods and strengthening of bacteriophage activities. Bacteriophage adsorption to host cells initiates the process of infection. The rotating magnetic field (RMF) is a promising biotechnological method for process intensification, especially for the intensification of micromixing and mass transfer. This study evaluates the use of RMF to enhance the infection process by influencing bacteriophage adsorption rate. The RMF exposition decreased the t50 and t75 of bacteriophages T4 on Escherichia coli cells and vb_SauM_A phages on Staphylococcus aureus cells. The T4 phage adsorption rate increased from 3.13 × 10-9 mL × min-1 to 1.64 × 10-8 mL × min-1. The adsorption rate of vb_SauM_A phages exposed to RMF increased from 4.94 × 10-9 mL × min-1 to 7.34 × 10-9 mL × min-1. Additionally, the phage T4 zeta potential changed under RMF from -11.1 ± 0.49 mV to -7.66 ± 0.29 for unexposed and RMF-exposed bacteriophages, respectively.

The Influence of Hydrodynamic Conditions in a Laboratory-Scale Bioreactor on Pseudomonas aeruginosa Metabolite Production.

Hydrodynamic conditions are critical in bioprocessing because they influence oxygen availability for cultured cells. Processes in typical laboratory bioreactors need optimization of these conditions using mixing and aeration control to obtain high production of the desired bioproduct. It could be done by experiments supported by computational fluid dynamics (CFD) modeling. In this work, we characterized parameters such as mixing time, power consumption and mass transfer in a 2 L bioreactor. Based on the obtained results, we chose a set of nine process parameters to test the hydrodynamic impact on a selected bioprocess (mixing in the range of 0-160 rpm and aeration in the range of 0-250 ccm). Therefore, we conducted experiments with P. aeruginosa culture and assessed how various hydrodynamic conditions influenced biomass, pyocyanin and rhamnolipid production. We found that a relatively high mass transfer of oxygen (kLa = 0.0013 s-1) connected with intensive mixing (160 rpm) leads to the highest output of pyocyanin production. In contrast, rhamnolipid production reached maximal efficiency under moderate oxygen mass transfer (kLa = 0.0005 s-1) and less intense mixing (in the range of 0-60 rpm). The results indicate that manipulating hydrodynamics inside the bioreactor allows control of the process and may lead to a change in the metabolites produced by bacterial cells.

Preparation of Komagataeibacter xylinus Inoculum for Bacterial Cellulose Biosynthesis Using Magnetically Assisted External-Loop Airlift Bioreactor.

The aim of this study was to demonstrate the applicability of a novel magnetically assisted external-loop airlift bioreactor (EL-ALB), equipped with rotating magnetic field (RMF) generators for the preparation of Komagataeibacterxylinus inoculum during three-cycle repeated fed-batch cultures, further used for bacterial cellulose (BC) production. The fermentation carried out in the RMF-assisted EL-ALB allowed to obtain an inoculum of more than 200× higher cellular density compared to classical methods of inoculum preparation. The inoculum obtained in the RMF-assisted EL-ALB was characterized by a high and stable metabolic activity during repeated batch fermentation process. The application of the RMF-assisted EL-ALB for K. xylinus inoculum production did not induce the formation of cellulose-deficient mutants. It was also confirmed that the ability of K. xylinus to produce BC was at the same level (7.26 g/L of dry mass), regardless of inoculum age. Additionally, the BC obtained from the inoculum produced in the RMF-assisted EL-ALB was characterized by reproducible water-related properties, mechanical strength, nano-fibrillar structure and total crystallinity index. The lack of any negative impact of inoculum preparation method using RMF-assisted EL-ALB on BC properties is of paramount value for its future applications, including use as a biomaterial in tissue engineering, wound healing, and drug delivery, where especially BC liquid capacity, nanostructure, crystallinity, and mechanical properties play essential roles.

Fabrication of Paper Sheets Coatings Based on Chitosan/Bacterial Nanocellulose/ZnO with Enhanced Antibacterial and Mechanical Properties.

Here, we designed paper sheets coated with chitosan, bacterial cellulose (nanofibers), and ZnO with boosted antibacterial and mechanical activity. We investigated the compositions, with ZnO exhibiting two different sizes/shapes: (1) rods and (2) irregular sphere-like particles. The proposed processing of bacterial cellulose resulted in the formation of nanofibers. Antimicrobial behavior was tested using E. coli ATCC® 25922™ following the ASTM E2149-13a standard. The mechanical properties of the paper sheets were measured by comparing tearing resistance, tensile strength, and bursting strength according to the ISO 5270 standard. The results showed an increased antibacterial response (assigned to the combination of chitosan and ZnO, independent of its shape and size) and boosted mechanical properties. Therefore, the proposed composition is an interesting multifunctional mixture for coatings in food packaging applications.

Transdermal Delivery Systems for Ibuprofen and Ibuprofen Modified with Amino Acids Alkyl Esters Based on Bacterial Cellulose.

The potential of bacterial cellulose as a carrier for the transport of ibuprofen (a typical example of non-steroidal anti-inflammatory drugs) through the skin was investigated. Ibuprofen and its amino acid ester salts-loaded BC membranes were prepared through a simple methodology and characterized in terms of structure and morphology. Two salts of amino acid isopropyl esters were used in the research, namely L-valine isopropyl ester ibuprofenate ([ValOiPr][IBU]) and L-leucine isopropyl ester ibuprofenate ([LeuOiPr][IBU]). [LeuOiPr][IBU] is a new compound; therefore, it has been fully characterized and its identity confirmed. For all membranes obtained the surface morphology, tensile mechanical properties, active compound dissolution assays, and permeation and skin accumulation studies of API (active pharmaceutical ingredient) were determined. The obtained membranes were very homogeneous. In vitro diffusion studies with Franz cells were conducted using pig epidermal membranes, and showed that the incorporation of ibuprofen in BC membranes provided lower permeation rates to those obtained with amino acids ester salts of ibuprofen. This release profile together with the ease of application and the simple preparation and assembly of the drug-loaded membranes indicates the enormous potentialities of using BC membranes for transdermal application of ibuprofen in the form of amino acid ester salts.

Bacterial Cellulose Membrane Containing Epilobium angustifolium L. Extract as a Promising Material for the Topical Delivery of Antioxidants to the Skin.

Bacterial cellulose membranes (BCs) are becoming useful as a drug delivery system to the skin. However, there are very few reports on their application of plant substances to the skin. Komagataeibacter xylinus was used for the production of bacterial cellulose (BC). The BC containing 5% and 10% ethanolic extract of Epilobium angustifolium (FEE) (BC-5%FEE and BC-10%FEE, respectively) were prepared. Their mechanical, structural, and antioxidant properties, as well as phenolic acid content, were evaluated. The bioavailability of BC-FESs using mouse L929 fibroblasts as model cells was tested. Moreover, In Vitro penetration through the pigskin of the selected phenolic acids contained in FEE and their accumulation in the skin after topical application of BC-FEEs was examined. The BC-FEEs were characterized by antioxidant activity. The BC-5% FEE showed relatively low toxicity to healthy mouse fibroblasts. Gallic acid (GA), chlorogenic acid (ChA), 3,4-dihydroxybenzoic acid (3,4-DHB), 4-hydroxybenzoic acid (4-HB), 3-hydroxybenzoic acid (3-HB), and caffeic acid (CA) found in FEE were also identified in the membranes. After topical application of the membranes to the pigskin penetration of some phenolic acid and other antioxidants through the skin as well as their accumulation in the skin was observed. The bacterial cellulose membrane loaded by plant extract may be an interesting solution for topical antioxidant delivery to the skin.

Modulation of Cellular Response to Different Parameters of the Rotating Magnetic Field (RMF)-An In Vitro Wound Healing Study.

Since the effect of MFs (magnetic fields) on various biological systems has been studied, different results have been obtained from an insignificant effect of weak MFs on the disruption of the circadian clock system. On the other hand, magnetic fields, electromagnetic fields, or electric fields are used in medicine. The presented study was conducted to determine whether a low-frequency RMF (rotating magnetic field) with different field parameters could evoke the cellular response in vitro and is possible to modulate the cellular response. The cellular metabolic activity, ROS and Ca2+ concentration levels, wound healing assay, and gene expression analyses were conducted to evaluate the effect of RMF. It was shown that different values of magnetic induction (B) and frequency (f) of RMF evoke a different response of cells, e.g., increase in the general metabolic activity may be associated with the increasing of ROS levels. The lower intracellular Ca2+ concentration (for 50 Hz) evoked the inability of cells to wound closure. It can be stated that the subtle balance in the ROS level is crucial in the wound for the effective healing process, and it is possible to modulate the cellular response to the RMF in the context of an in vitro wound healing.

Single Mathematical Parameter for Evaluation of the Microorganisms' Growth as the Objective Function in the Optimization by the DOE Techniques.

The cultivation of bacteria sets a ground for studying biological processes in many scientific disciplines. The development of the bacterial population is commonly described with three factors that can be used to evaluate culture conditions. However, selecting only one of them for the optimization protocol is rather problematic and may lead to unintended errors. Therefore, we proposed a novel mathematical approach to obtain a single factor that could be used as the objective function to evaluate the whole growth dynamic and support the optimization of the biomass production process. The sigmoidal-shape curve, which is the commonly used function to plot the amount of biomass versus time, was the base for the mathematical analysis. The key process parameters, such as maximal specific growth rate and lag-phase duration were established with the use of mathematical coefficients of the model curve and combined to create the single growth parameter. Moreover, this parameter was used for the exemplary optimization of the cultivation conditions of Klebsiella pneumoniae that was cultured to be further used in the production of lytic bacteriophages. The proposed growth parameter was successfully validated and used to calculate the optimal process temperature of the selected bacterial strain. The obtained results indicated that the proposed mathematical approach could be effortlessly adapted for a precise evaluation of growth curves.

Evaluation of usefulness of 2DCorr technique in assessing physicochemical properties of bacterial cellulose.

The main aim of the current study was to evaluate the usefulness of the two-dimensional correlation analysis (2DCorr) technique in assessing the physicochemical properties of bacterial cellulose (BC) depending on the synthesis time and Komagataeibacter xylinus strain used for its production. It was demonstrated that 2DCorr analysis allows to characterize substantial changes in the structure of BC at the molecular level occurring during its synthesis, which are difficult to determine using ATR-FTIR spectral analysis. Furthermore, it was found that 2DCorr analysis allows to identify the differences in the structure and the properties of BC according to the bacterial strain used for its synthesis. It was concluded that 2DCorr analysis of the ATR-FTIR spectra may be a useful tool for controlling BC production in order to obtain material with the desired properties.

Increased water content in bacterial cellulose synthesized under rotating magnetic fields.

The current study describes properties of bacterial cellulose (BC) obtained from Komagataeibacter xylinus cultures exposed to the rotating magnetic field (RMF) of 50 Hz frequency and magnetic induction of 34 mT for controlled time during 6 days of cultivation. The experiments were carried out in the customized RMF exposure system adapted for biological studies. The obtained BC displayed an altered micro-structure, degree of porosity, and water-related parameters in comparison to the non-treated, control BC samples. The observed effects were correlated to the duration and the time of magnetic exposure during K. xylinus cultivation. The most preferred properties in terms of water-related properties were found for BC obtained in the setting, where RMF generator was switched off for the first 72 h of cultivation and switched on for the next 72 h. The described method of BC synthesis may be of special interest for the production of absorbent, antimicrobial-soaked dressings and carrier supports for the immobilization of microorganisms and proteins.

Modification of bacterial cellulose through exposure to the rotating magnetic field.

The aim of the study was to assess the influence of rotating magnetic field (RMF) on production rate and quality parameters of bacterial cellulose synthetized by Glucanacetobacter xylinus. Bacterial cultures were exposed to RMF (frequency f=50Hz, magnetic induction B=34mT) for 72h at 28°C. The study revealed that cellulose obtained under RMF influence displayed higher water absorption, lower density and less interassociated microfibrils comparing to unexposed control. The application of RMF significantly increased the amount of obtained wet cellulose pellicles but decreased the weight and thickness of dry cellulose. Summarizing, the exposure of cellulose-synthesizing G. xylinus to RMF alters cellulose biogenesis and may offer a new biotechnological tool to control this process. As RMF-modified cellulose displays better absorbing properties comparing to non-modified cellulose, our finding, if developed, may find application in the production of dressings for highly exudative wounds.