INMdok (Leibniz Institute for New Materials)
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Engineered microbial living matter for diagnostics, prevention, and therapy
Full text linkLiving therapeutic and diagnostic materials based on engineered microorganisms are emerging as a novel approach with the perspective of providing patient-tailored, sustainable, and cost-effective healthcare solutions. In this review, we focus on recent advances in using genetically or chemically engineered microorganisms as living diagnostics, therapeutics, and as a means of prevention for various diseases. We also highlight the applications of living therapeutics for acute and chronic diseases, and the role of micro/macro-encapsulation of the engineered microorganisms. We further showcase the current success of engineered living therapeutics in clinical trials and discuss challenges and future trends in the field
A biocompatible polylactide-ε-caprolactone polymer coated with poly(hexamethylene biguanide) displays antibacterial properties against slime-producing S. epidermidis
Full text linkIntroduction: produced by renewable resources, biodegradable polymers with their competitive mechanical properties, thermal stability and biocompatibility are important alternatives to other synthetic materials for use in medical devices, i.e. endotracheal suction catheters. However, infected catheters may lead to nosocomial infections, such as lower respiratory tract infections, with mechanical ventilation being a major risk for these. Antimicrobially coated endotracheal suction catheters may be one measure to reduce this risk. Methods: two procedures using ethanol and sodium hydroxide were tested to immobilize poly(hexamethylene biguanide) (PHMB) to polylactide-ε-caprolactone (PLA-ε-CL). The cytocompatibility of the coating was verified via the MTT assay and cytokine analysis in a cell monolayer and in a 3D mucosa model. The antimicrobial efficacy was tested using S. epidermidis; after this bacterial contamination and the adherence and viability of cells were tested. Chemical surface analysis has been performed with pristine and PHMB-coated specimens by means of infrared spectroscopy (ATR-FTIR). Results: with both applied coating procedures, PHMB could be immobilized onto the PLA-ε-CL surface. The biocompatibility of PLA-ε-CL was not impaired by the PHMB coating. IL-1α was slightly but significantly increased. Reduction of S. epidermidis was about 4 lg-levels after 6 h of incubation. Contamination of the surface prior to cell culture did not impair the adherence of the cells. Conclusion: we demonstrated that PLA-ε-CL coated with PHMB has good biocompatible properties with antimicrobial activity thus revealing the polymer to be a suitable material for the development of medical devices that are able to prevent bacterial contaminations and infections
Data Management Plan in RSpace: 'One tool less' to simplify research workflow
Full text linkThe effective management of research data begins with creating a data management plan (DMP) for each research project which describes how the research data is handled throughout the research life cycle. However, the importance of proper handling of research data is not realized among researchers as research data management (RDM) is not embedded in their everyday practice. Researchers often get overwhelmed by any additional tools or infrastructure introduced to their research workflows. Therefore, to streamline the research process, we propose a “one less tool” strategy, integrating DMP template into our electronic laboratory notebook (ELN) “RSpace” which helps researchers use existing institutional services and infrastructure instead of adopting an additional platform for creating DMP. This strategy ensures that researchers can focus more on their work while maintaining efficiency in data management. Currently, we are in a trail phase to evaluate the effectiveness of this integration. We assume that researchers are more likely to create their DMPs when the process is integrated in their ELN, reducing the need for an extra effort
Effects of formaldehyde on YAP and NF-κB signaling pathways in mammalian cells
Full text linkFormaldehyde is the smallest existing aldehyde, a highly reactive color less gas at room temperature and ubiquitously present in our atmosphere. Because of its reactivity leading to the crosslinking of macromolecules like proteins, it is widely used in industrial applications, but also in cell biology in order to preserve cells and tissues for further analysis. In this work, we show that formaldehyde releasing solutions commonly used for fixation of cells, can diffuse via the gas phase to the neighboring well and influence signaling processes in the therein cultured cells. To analyze this effect, we utilized a stable reporter cell line for YAP signaling or a gene expression-based reporter for activation of the NF-κB pathway. We could show that next to formaldehyde, also glutaraldehyde and acetaldehyde were able to activate those signaling pathways. Additionally, especially the stable reporter cell line based on YAP signaling can also be used as sensor for bioavailable formaldehyde, being highly sensitive, easy to use, and reversible. The observed impact of formaldehyde on cellular signaling underscores the need for careful planning of experimental protocols and emphasizes the importance of implementing proper controls when utilizing this reagent in cellular signaling analyses
Reversibly Charge-Switching Polyzwitterionic/Polycationic Coatings for Biomedical Applications: Optimizing the Molecular Structure for Improved Stability
Full text linkMaterials that can be switched between a polycationic/antimicrobial and a polyzwitterionic/protein-repellent state have important applications, e.g., as biofilm-reducing coatings in medical devices. However, the lack of stability under storage and application conditions so far restricts the lifetime and efficiency of such materials. In this work, a polynorbornene-based polycarboxybetaine with an optimized molecular structure for improved hydrolytic stability is presented. The polymer is fully characterized on the molecular level. Surface-attached polymer networks are obtained by spin-coating and UV cross-linking. These coatings are highly uniform and demonstrate charge-switching in zeta-potential studies. Storage stability in the dry state, as well as in aqueous systems at pH 4.5 and 7.4 for 28 days, is demonstrated. At pH 8, hydrolytic degradation is observed. Overall, the materials are substantially more stable than the corresponding ester-based systems
Optimizing the composition of bioactive coatings to support toluene removal
Full text linkThe potential of bioactive coatings as an innovative biotechnology to overcome the mass-transfer limitations of conventional technologies when treating air pollutants, especially hydrophobic volatile organic compounds, was herein assessed. Bioactive coatings consist of active microorganisms entrapped in a polymer matrix, which needs to be porous to facilitate an effective gas pollutant exchange. To increase porosity, two additives, sucrose and glycerol mixtures (Suc/Gly) and halloysite nanotubes (HNTs), were included in the bioactive coatings at two concentration levels. The toluene removals of the different bioactive coatings were studied in batch mode at low (∼300 mg m−3) and high (∼3000 mg m−3) toluene concentrations. Overall, low HNTs concentration coatings supported optimum toluene removals (>95 %), comparable to biofilm controls at both toluene concentrations. High HNTs concentration coatings and low Suc/Gly concentration coatings achieved toluene removals over 95 % after 7 toluene injections at low toluene concentration. At high toluene concentrations, these coatings eventually outperformed the biofilm controls. High Suc/Gly concentration coatings supported a limited toluene removal (4 and 1 injection at low and high toluene concentrations, respectively), attributed to a preferential consumption of sucrose over toluene. These findings were corroborated by ESEM/conventional SEM imaging, revealing porosity in the HNTs bioactive coatings, visible at both the surface and internal levels. On the contrary, more homogeneous surfaces were observed in the Suc/Gly bioactive coatings, where total polymer coalescence was partially hindered by the addition of Suc/Gly. These results paved the way towards the implementation of bioactive coating in larger bioreactors for real-life air purification
Encapsulation-enhanced genetic switches in lactobacilli
No full textLactiplantibacillus plantarumis known for its potential in healthcare, food production, and environmental biotechnology. However, its broader utility is constrained by a limited genetic toolbox, particularly lacking robust genetic switches for inducible gene expression. Addressing this gap, we developed a novel genetic switch forL. plantarumbased on a strong bacteriophage-derived promoter and the food-grade inducer, cumate. However, the switch was susceptible to leaky expression in the late log phase of bacterial growth, which was correlated to a reduction in the culture pH. This leakiness was partially resolved by regulating culture conditions (temperature and nutrients) to limit growth below a certain bacterial density. More interestingly, leaky expression could be stably suppressed by encapsulating the bacteria in alginate as an engineered living material. This physically restricted growth and limited the pHdrop, thereby enhancing the switch performance. The possibilities to regulate protein secretion over several days, reversibly switch protein production, and establish dual functionalities by co-encapsulating strains with different switches were demonstrated. Thus, for the first time, we show a material-based strategy to enhance the performance of a genetic switch in bacteria. This strategy facilitates the development ofL. plantarumfor advanced applications in biotechnology, pharmaceutics, and living therapeutics
Sensing relative humidity with a fluorescent seed-like biodegradable flier
Full text linkPlant-inspired soft robots enable distributed environmental monitoring. Fliers, i.e. soft robots that are carried passively by the wind, can be effectively deployed and cover large areas and distances. State-of-the-art fliers for humidity sensing are largely composed of electronic components, which increase cost and generate electronic waste. Here, we introduce self-deployable and biodegradable fliers inspired by natural Ailanthus altissima seeds. These artificial fliers are composed of fluorescent, cellulose-based composites with sensing capabilities. The material is shaped into artificial seeds using scalable 3D extrusion processing. Red-emitting Mn2+-doped Er3+, Yb3+:NaYF4 nanoparticles in the composite provide a strong optical emission upon excitation at 980 nm wavelength. The cellulose matrix absorbs water, which quenches the intensity of fluorescence of the nanoparticles. Increasing humidity thus changes the color of the fluorescence emission from red to green. We used ratiometric sensing to detect the humidity of the surroundings
Multilayered trabecular meshwork for dynamic in vitro studies in glaucoma research
Full text linkGlaucoma, an eye disease causing incremental vision loss, currently has no cure. Its primary cause is the malfunction of the trabecular meshwork (TM), a multilayered tissue in the eye responsible for draining aqueous humor (AH) from the anterior chamber. TM clogging increases outflow resistance, elevates intraocular pressure (IOP), and damages optic nerves, leading to irreversible blindness. Existing in vitro TM models are suboptimal, as they lack the hierarchical structure of the TM. This article introduces a dynamic in vitro TM model, featuring a multilayered scaffold architecture 3D printed via melt electrowriting (MEW), and integrated with a flow system that enables continuous pressure monitoring during perfusion at native flow rates. Printed scaffolds supported the growth of primary adult human TM cells that grew on top and between the fibers. Cellularized scaffolds were tested under static and dynamic conditions. Over 3–5 days, pressure monitoring showed increased outflow resistance due to cell proliferation. Proteomic analysis revealed distinct changes in protein expression related to protein synthesis and respiration of cells grown under flow. Lat-B administration resulted in decreased pressure values and depolymerized actin filaments. These findings suggest that the proposed model is a promising alternative for in vitro glaucoma drug testing
Direct Ink Writing of Carbon‐Based Electrofluids for Soft Electrical Component Manufacturing
Full text linkAbstractSoft electrical components are highly demanded for human‐machine interaction devices. “Electrofluids” (EFs), which are suspensions of electrically conductive filler particles in non‐conductive solvents, are proposed as promising sensors and conductive materials since they can flow and retain electrical conductivity. As they remain liquid in working conditions, encapsulation and manufacturing of complex patterns remain as a challenge but would enable a wider variety of applications. Direct ink writing (DIW) is proposed here as a method to manufacture carbon‐based EFs. Simple shear flow and Fourier‐transform (FT) rheology are performed to evaluate the printability of EFs containing different concentrations of Carbon Black and Graphene Powder by DIW. Electrofluids exhibited three important characteristics to be manufactured via DIW: yield stress behavior (confirmed by flow curves), high brittleness, and a fast mechanical recovery within a range of 15 s. Printability maps are created to distinguish printable and non‐printable EFs. Printable EFs are used to manufacture complex patterns. As a proof of the great potential of the EFs and DIW combination, a comparison between simple and multiline strain gauges showed an enhancement in the sensitivity of EFs as strain sensors by almost 800%