Introduction:

Bioengineering plays a crucial role in developing advanced biomedical devices and interfaces that integrate biological systems. One major challenge in the development of cell-on-a-chip interfaces is preventing bacterial contamination while maintaining cellular compatibility. Low-fouling (super-)hydrophilic zwitterionic polymer materials have emerged as potential biomedical materials and bio-functional coatings. Here, we report a novel terpolymer nano-brush coating that effectively suppresses undesired biomolecular fouling and biofilm formation while providing sufficient molecular functionalization capacity..

Methods

A terpolymer brush nanolayer composed of carboxybetaine methacrylamide (CBMAA), N-(2-hydroxypropyl) methacrylamide (HPMAA), and sulfobetaine methacrylamide (SBMAA) was synthesized on glass and gold-coated glass substrates using the ATRP method. The chemical structure, thickness, surface zeta potential, fouling resistance, and wettability of this coating were analyzed using XPS, FT-IRRAS, spectroscopic ellipsometry, electrokinetic analyzer, SPR, and water contact angle measurements. Bacterial and cell adhesion studies were conducted on bare and RGD-functionalized terpolymer brushes.

Results

In vitro analyses confirmed the coating’s exceptional resistance against Staphylococcus epidermidis and Pseudomonas aeruginosa while enhancing macrophage mobility compared to uncoated glass, likely due to the coating’s highly hydrated nature and low protein adsorption. RGD-functionalized terpolymer coatings were found to be non-cytotoxic for SaOS-2 osteosarcoma cells, promoting cell adhesion and spreading without significantly increasing bacterial adhesion or protein adsorption. These results demonstrate that bio-functional terpolymer nano-brushes provide a dual benefit of bacterial resistance and cellular compatibility, making them highly prospective for cell-on-a-chip applications.

Discussion

This study discusses the importance of combining antifouling efficacy, enhanced cellular interaction, and non-cytotoxicity when developing tailored materials for next-generation cell-on-a-chip interfaces. Future research will focus on scaling up the synthesis process and exploring the long-term stability and biocompatibility of these nano-brushes in various bioengineering applications.

Introduction: In industry, α-ketoglutaric acid (KGA) is produced by chemical synthesis. A more promising approach is the microbiological synthesis of KGA using the yeast Yarrowia lipolytica. This approach allows for the production of a high-quality product, which is of significant interest to the food and pharmaceutical industries. Ethyl alcohol, glycerol, vegetable oils, and n-alkanes can be employed as the carbon sources for KGA production by Y. lipolytica. Despite the extensive body of literature on the biosynthesis of KGA by Y. lipolytica, there is no information on scale-up experiments. Concurrently, the approbation of the technology for the synthesis of KGA in pilot bioreactors is a prerequisite for the transition to industrial production.

Methods: The yeast strain Y. lipolytica VKM Y-2412 was cultivated in a 1000-liter SGI bioreactor (Setric Genie Industriel, Toulouse, France) with 400 liters of Reeder medium at 30°C.

Results and Discussion: For the first time, it was demonstrated that it is feasible to produce KGA under pilot conditions. Additionally, technological approaches to realisation of the process were developed. The producer demonstrated optimal growth at a pH of 4.5 and a dissolved oxygen (pO₂) level of 25% saturation during the first two days. Following a two-day period, the production of KGA commenced. Throughout the acid production process, the pH and aeration levels were maintained at 3.5 and 50-55%, respectively. Additionally, rapeseed oil was added at a concentration of 20 g/L when the pO₂ value increased by 10% above the stable level. After 140 hours of cultivation, Y. lipolytica VKM Y-2412 produced 42 g/L of KGA, representing 30% of the rapeseed oil consumed.

Conclusions: This research constituted the first attempt to produce KGA in a large-scale bioreactor. Further research could be conducted to optimize the cultivation conditions at the pilot scale in order to increase the product yield.

Biological structures engage in direct environmental interaction and acquire knowledge through multi sensory feedback from sensory input stimulants that influence the inner neuron structures that are formed. We describe a robotic system that uses multi sensory learning to handle things, taking influence from biological principles like the exploring and processing of sensory information, which ultimately lead to behavior learning. The robot can communicate smartly with its surrounding environment due to a small-scale biological neuromorphic prosthetic circuit that locally combines and interprets multi modal sensory input stimulants in an adaptable manner. Through the use of low-voltage organically neurological devices with a synapse capacity to handle sensory input stimulants in real-time, multi-sensory-associated linkages are formed, which eventually lead to behavioral training and the robot learning to avoid possibly hazardous things. A key component of the neuromorphic circuit's functionality is the employment of functioning components, such as organically semi-conducting compounds of polymer, which replicate bio-inspired features including dendritic summary, the plasticity of synapses, and neural computation. The monolithic polymeric electronics that are low-power, locally unified, and on a tiny scale can do this. Furthermore, the idea of handling sensory data of variable complexity and multifaceted communication can be expanded into several branches by virtue of the neuromorphic factor circuit's modular-like construction. This robotic device provides a concrete illustration of how localized organic neuromorphic factor circuits combined with bio-influenced principles might result in the creation of extremely adaptable, smart, and efficient systems for practical use.

Introduction:

Cell analysis is important in the diagnosis of several diseases, as the dielectric and morphological properties of the cells can change when diseased. In this context, malaria is one of the most threatening diseases affecting red blood cells (RBCs), leading to their destruction. High-frequency impedance spectroscopy (HFIS) with nanoelectrode arrays has the potential to overcome the Debye screening effect of the electrolyte and surpass the cellular membrane, thus allowing the study of the cell internal structure. In this work, we investigate, by simulation, the potential of the NXP CMOS nanoelectrode array biosensing platform in Widdershoven2018 to study the internal structure of the cells.

Methods:

Simulations were carried out in COMSOL Multiphysics ®, modelling a healthy RBC discocyte with an overall volume of 86 fL. The malaria parasite within the cell was modelled at three different stages of infection: 6 hpi (hours post infection), 12 hpi, and 18hpi. The simulation medium consisted of a physiological solution that does not affect the cell’s properties. All simulation parameters were taken from Honrado2018.

Results and Discussion:

The simulations were performed with an RBC positioned in the centre of the array, obtaining ∆C as the difference in capacitance with and without the cell. The capacitance profiles show a change in the ∆C spectrum above 1 MHz, due to the malaria parasite. Analyzing the difference in capacitance between uninfected and infected RBCs at high frequencies, the variation reaches valued as high as 100 aF, measurable with the existing NXP nanoelectrode array platform. This provides a framework for distinguishing between healthy cells and the ones with intracellular inclusions.

Conclusions:

The potential of HFIS analysis with a CMOS nanoelectrode array platform was explored in this work, suggesting the possibility of a label-free and non-invasive method to study and explore the internal structure of a cell.

The development of novel scaffolds for bone tissue engineering offers a promising approach to promote bone regeneration. This study aimed to develop a support structure using electrospinning, combining PCL fibers and bioglass powder for bone regeneration. The solution contained 15% PCL/1% bioglass. Tests analyzed the structure's chemical and biological properties. Support structures were produced using electrospinning, and fiber diameter was measured using ImageJ on scanning electron microscopy (SEM) images. Chemical analysis used Energy-Dispersive X-ray Spectroscopy (EDS) and Fourier Transform Infrared Spectroscopy (FTIR) spectroscopy. Cytotoxicity was assessed through a lactate dehydrogenase (LDH) release test on tooth stem cells. Cell viability was analyzed using the 3-4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, and alkaline phosphatase activity evaluated osteogenic differentiation. The PCL/BC scaffolds exhibited elongated, granule-free fibers, with a mean diameter and standard deviation of 3.8 ± 1.4 µm. EDS revealed BC constituents (carbon, sodium, calcium, silicon, phosphorus and oxygen), indicating the incorporation of BC powder into the fibers. FTIR spectroscopy showed C=O and OC-O interactions. The contact angle indicated hydrophobicity in PCL, PCL/BC, but after two minutes, the PCL/BC scaffold exhibited the expected hydrophilicity. PCL/BC scaffolds did not show cytotoxicity based on LDH release. In osteogenic media, cell viability was higher in PCL/BC compared to PCL and standard well plates after six days, while lactate dehydrogenase levels remained similar across the plate, PCL, and PCL/BC groups, showing higher activity than the undifferentiated control. This study demonstrated that electrospun PCL/BC structures with 15% PCL and 1% BC had advantageous properties, including long fibers and increased hydrophilicity. These structures showed no cytotoxicity and high cell viability, suggesting their potential for bone regeneration.

Support: The Office of Naval Research Global (ONRG Award N62909-21-1-2026); National Institute of Science and Technology for Regenerative Medicine (INCT-Regenera); Stem Cell Research Institute (IPCT).

Introduction: The execution of complex motor actions involving processes such as planning, decision-making, and execution entails a certain cognitive workload (CWL), engaging the involvement of multiple brain areas and processes related to the coordinated activation of muscles. Factors such as context, previous experience, stimulus complexity, and required precision, among others, can establish basal cortical patterns from which those related to the specific motor task are generated. Here, we propose a study to characterize cortical patterns before and after the execution of musical motor tasks.

Methods: Ten subjects participated in an experimental involving the unilateral execution of musical chord on an electronic piano. Nine musical chord (C, D, E₇, F, F_#m, G, A, A_m, and B) were presented alternately and sequentially to the subjects on a 21-inch monitor. Throughout the experiment, EEG signals were recorded from 256 channels. Cortical activity was analyzed based on the spectral energy in different bands and patterns of connectivity evoked before and after the motor executions.

Results and Discussion: Event-related desynchronizations and connectivity patterns have shown differential characteristics before and after musical tasks. Moreover, the presentation of musical chords triggered cortical patterns different from those evoked by the instruction of a simple motor action (pressing a piano key).

Conclusions: With this study, we characterized the cortical dynamics evoked by the performance of musical chords. Likewise, we concluded that it would be possible to determine the cognitive loads demanded on subjects by musical instructions measuring EEG signals.

Tissue engineering and regenerative medicine have new meanings. Three-dimensional bioprinting has become one of the most advanced and useful innovations that allows the creation of personalized macroscopic and microscopic constructs at different scales that match a patient’s anatomy. Generally, the process of 3D bioprinting consists of several steps, namely pre-bioprinting, cell and bioink preparation, the bioprinting process, and post-bioprinting/applications. Intensive research efforts are currently underway to develop highly printable and biocompatible materials. Among the variety of bioprinting materials (i.e., biomaterial inks), naturally derived hydrogels have attracted great interest due to their beneficial properties in terms of biocompatibility, cost effectiveness, and biodegradability. Among them are cellulose, chitosan, and lignin. Cellulose is the most abundant biopolymer in nature; it has various advantages over others, such as good mechanical and barrier properties. Moreover, chitosan biopolymer constitutes a promising candidate for the preparation of hydrogels for application in this field due to its beneficial properties, such as its antimicrobial activity and structural resemblance to natural glycosaminoglycans. Furthermore, due to the non-cytotoxicity, biocompatibility, biodegradability, mechanical strength, and reactivity of the lignin biopolymer, it has been considered an excellent candidate to manufacture hydrogels for 3D bioprinting applications. The capacity of 3D bioprinting of these biopolymer-based hydrogels has been demonstrated in the regeneration of different damaged tissues, including cartilage, bone, muscle, skin, blood vessels, and other biological tissues. In this study, we provide a comprehensive review of the formulation and use of three functional biomaterials as ink-based hydrogels. Cellulose, chitosan, and lignin are comprehensively discussed, and an examination of the status of the biomedical application of these biopolymer-based hydrogels for 3D bioprinting is then provided.

Introduction: Skin wounds present a considerable challenge, impacting millions globally. This research aimed to create a bioink utilizing lyophilized rat decellularized skin (DS) for 3D bioprinting to improve skin regeneration. Methods: Rat skin was subjected to decellularization for 5 days. A comparative analysis of genomic DNA quantification and histological staining was performed between native and decellularized skin. The tissue was freeze-dried and combined with alginate and gelatin to formulate bioinks with concentrations of 1.5% and 3% DS, 3% or 4% alginate, and 7% gelatin. Rheological evaluations, including swelling, printability, and degradation over a four-week period, were conducted. Hydrogel SEM images were obtained using a scanning electron microscope. Cell viability and proliferation were assessed using the Live/Dead assay.

Results: The hydrogel demonstrated good shear-thinning behavior and maintained its viscosity across different concentrations. The degradation rate was 59,2% in one month. Swelling was 3783% after one month. Only the bioink with 1.5% DS, 3% alginate, and 7% gelatin preserved structural integrity for four weeks and was chosen for further examination. Furthermore, the bioink showed a low tangent delta, decreasing printing-related stress and subsequent cell death. SEM images revealed a porous three-dimensional structure. The Live/Dead assay indicated higher cell viability (65%) compared to the control seven days post-bioprinting.

Conclusion: The biomaterial showed good mechanical properties and, after bioprinting, supported cell proliferation, indicating its potential as a promising alternative for skin wound regeneration.

Funding: The Office of Naval Research Global (ONRG Award N62909-21-1-2026), The National Institute of Science and Technology for Regenerative Medicine (INCT-Regenera) and The Stem Cell Research Institute (IPCT).

Plant biotechnology has presented the scientific world with a plethora of challenges and possibilities for decades. One of the most meticulously studied features of plant biomass is callus—unorganised, undifferentiated cells with the ability to heal wounded plant tissues or even regenerate whole organisms. Nicotiana tabacum Bright Yellow-2 (BY-2) is one of the most commonly used callus cell lines in research. These cells are able to grow as a suspension, and given their eukaryotic nature, with genetic modifications, they are able to produce mammalian proteins. This feature makes them extremely interesting, as they are likely to be used in biosimilar product manufacturing in the future. They are, however, prone to shear stress, which is the reason a bubble–free, wave–agitated bioreactor was used in this study.

ReadyToProcess WAVE 25 was used, along with a 2 L culture vessel, and cultures were performed, monitoring the effect of changing the angle (α) and frequency (ω) of the oscillations on the BY-2 cells. The dissolved oxygen level was monitored, and the biomass concentration was measured. Moreover, Western blotting was conducted to determine the presence of a protein product.

It was determined that the highest product activity and biomass proliferation was obtained at 0–2 days at α = 6° and ω = 20 min^-1, 3–5 days at α = 8° and ω = 26 min^-1, and 6–10 days at α = 12° and ω = 30 min^-1.

In conclusion, a correlation between the mixing conditions and the process’s efficiency was observed. However, further research is needed to establish the exact nature of the observed effects.

Background: Utilising stem cells is a promising approach to treat chronic periodontitis. Limited data is available regarding the regenerative potential of osteoporotic periodontal ligament stem cells (OP-PDLSCs). The IGF axis including its binding proteins (IGFBPs) is involved in osteogenesis. IGFBP-4 is particularly inhibitory of IGF function in vitro. Hence, the aim of this project was to study the osteogenic differentiation capacity of OP-PDLSCs and assess the possible role of IGF axis, particularly IGFBP-4 and its protease (PAPP-A), in the process.

Methods: PDLSCs were characterised from healthy (H-PDLSCs) and osteoporotic donors (both n=3). To compare their osteogenic differentiation, cells were cultured in basal media (control) or osteogenic media (supplemented with 50µM L-ascorbic acid and 10µM dexamethasone). Differentiation was assessed at 2, 3 and 4 weeks using Alkaline Phosphatase (ALP) and Alizarin Red staining (ARS) assays. RT-qPCR was conducted to assess the expression of the osteogenic and IGF axis markers. ELISA was used to measure IGFBP-4 and PAPP-A proteins.

Results: The intensity of ALP staining under osteogenic conditions increased with time for H-PDLSCs but not in OP-PDLSCs (except at week 4). ARS quantification indicated increased mineral concentration under osteogenic conditions after 3 and 4 weeks in H-PDLSCs (average 0.041±0.033 mM) compared to OP-PDLSCs (average 0.011±0.004 mM). Lower gene expression of the osteogenic markers (ALPL, RUNX2, Osteocalcin and Col 1A1) was measured in OP-PDLSCs, particularly in osteogenic conditions. In OP-PDLSCs, the strongest trend for downregulation was observed for IGFBP1, under both culture conditions, whilst other molecules were more variable. IGFBP-4 protein showed slightly elevated levels in OP-PDLSCs while PAPP-A levels (IGFBP-4 protease) were below detection.

Conclusion: OP-PDLSCs showed lower osteogenic differentiation capacity and different pattern of IGF axis expression compared to healthy controls. Manipulating IGF axis may be a strategy to enhance periodontal regeneration in OP patients.