Biohybrid Systems Research Articles

Polymer Materials Synthesized through Cell-Mediated Polymerization Strategies for Regulation of Biological Functions

ConspectusAs essential components of living organisms, biomacromolecules construct cell scaffolds and regulate cell activities and biological functions through chemical transformations in biological systems. Inspired by the functional evolution in the formation of natural structures, in situ polymerization methods have been developed to create functional synthetic macromolecules inside or on the surface of living cells. Given the diversity of cell species and the complexity of biological pathways, selected strategies can be employed to control the synthesis of functional polymers that utilize the dynamic cellular microenvironment.In this Account, we summarize recent work in the field of designing cell-mediated in situ polymerization methods, with which we demonstrate their application prospects including tumor cell labeling and treatment, microbial photosynthetic efficiency regulation, and hydrogel generation. The purpose of these efforts is to design polymerization reactions in response to endogenous or exogenous stimuli and to describe the underlying response mechanisms. By reasonable design of molecular structures, in situ synthesized polymers in the cell microenvironment implement regulation of biological functions. For example, using specific redox activity combined with light irradiation, bacteria can mediate the generation of functional polymers as the encapsulating matrix or with antibacterial effects. Conjugated polymers synthesized on the microalgae surface expanded the spectral absorption and improved photosynthetic efficiency. Meanwhile, characteristics of the cellular microenvironment could initiate various polymerization reactions inside living cells, including oxidative thiol cross-linking, condensation polymerization, and free radical polymerization. These reactions can be selectively conducted with reactive species generated in tumor cells, and the resulting polymers showed prolonged retention inside cells for modulating cell behaviors. Further development of cell-mediated polymerization strategies would provide an innovative platform for research and applications of multifunctional biomaterials and engineered biohybrid systems.

The perceived effects of augmented trail sensing and mood recognition abilities in a human–fish biohybrid system

The use of technologies to enhance human and animal perception has been explored in pioneering research about artificial life and biohybrid systems. These attempts have revealed that augmented sensing abilities can emerge from new interactions between individuals within or across species. Nevertheless, the diverse effects of different augmented capabilities have been less examined and compared. In this work, we built a human–fish biohybrid system that enhanced the vision of the ornamental fish by projecting human participants onto the arena background. In contrast, human participants were equipped with a mixed-reality device, which visualized individual fish trails (representing situation-oriented perceptions) and emotions (representing communication-oriented perceptions). We investigated the impacts of the two enhanced perceptions on the human side and documented the perceived effects from three aspects. First, both augmented perceptions considerably increase participants’ attention toward ornamental fish, and the impact of emotion recognition is more potent than trail sense. Secondly, the frequency of human–fish interactions increases with the equipped perceptions. The mood recognition ability on the human side can indirectly promote the recorded positive mood of fish. Thirdly, most participants mentioned that they felt closer to those fish which had mood recognition ability, even if we added some mistakes in the accuracy of mood recognition. In contrast, the addition of trail sensing ability does not lead to a similar effect on the mental bond. These findings reveal several aspects of different perceived effects between the enhancements of communication-oriented and situation-oriented perceptions.

Emerging Trends in Nanomaterials for Photosynthetic Biohybrid Systems

Global warming and climate change are among the most immediate challenges confronting humans in the 21st century. Artificial photosynthesis represents a promising approach to mitigating the environmental crisis. Recently, people demonstrated that interfacing semiconductor, polymer, or metal-based nanomaterials with specific bacteria can generate built-in artificial photosynthetic systems, enabling solar-to-fuel conversion by forming a basic photosynthetic unit from a network of light-harvesting receptors, molecular water splitting and CO2, or proton reduction machinery. As a cutting-edge research direction, several strategies have been employed to create the artificial photosynthetic biohybrids. Notably, understanding of the molecular basis of these photosynthetic biohybrid systems is the key to improving the solar-to-chemical conversion efficiency. In the current review, we highlight the study of charge uptake channels in biohybrid artificial photosynthetic systems using various nanomaterials and microbes. We emphasize the importance of fully understanding the structures and operating mechanisms of these hybrid systems, as well as the criterion to select suitable microbes and photosensitized nanomaterials.

Fluidized or not fluidized? Biophysical characterization of biohybrid lipid/protein/polymer liposomes and their interaction with tetracaine

BackgroundNanomedicine and the pharmaceutical industry demand the investigation of new biomaterials to improve drug therapies. Combinations of lipids, proteins, and polymers represent innovative platforms for drug delivery. However, little is known about the interactions between such compounds and this knowledge is key to prepare successful drug delivery systems. MethodsBiophysical properties of biohybrid vesicles (BhVs) composed of phospholipids, proteins, and amphiphilic block copolymers, assembled without using organic solvents, were investigated by differential scanning calorimetry and dynamic light scattering. We studied four biohybrid systems; two of them included the effect of incorporating tetracaine. Thermal changes of phospholipids and proteins when interacting with the amphiphilic block copolymers and tetracaine were analyzed. ResultsLysozyme and the copolymers adsorb onto the lipid bilayer modifying the phase transition temperature, enthalpy change, and cooperativity. Dynamic light scattering investigations revealed relevant changes in the size and zeta potential of the BhVs. Interestingly, tetracaine, a membrane-active drug, can fluidize or rigidize BhVs. ConclusionsWe conclude that positively charged regions of lysozyme are necessary to incorporate the block copolymer chains into the lipid membrane, turning the bilayer into a more rigid system. Electrostatic properties and the hydrophilic-lipophilic balance are determinant for the stability of biohybrid membranes. General significanceThis investigation provides fundamental information associated with the performance of biohybrid drug delivery systems and can be of practical significance for designing more efficient drug nanocarriers.

Is biorobotics science? Some theoretical reflections

In this paper, we ask one fairly simple question: to what extent can biorobotics be sensibly qualified as science? The answer clearly depends on what ‘science’ means and whether what is actually done in biorobotics corresponds to this meaning. To respond to this question, we will deploy the distinction between science and so-called technoscience, and isolate different kinds of objects of inquiry in biorobotics research. Capitalising on the distinction between ‘proximal’ and ‘distal’ biorobotic hypotheses, we will argue that technoscientific biorobotic studies address proximal hypotheses, whilst scientific biorobotic studies address distal hypotheses. As a result, we argue that bioroboticians can be both considered as scientists and technoscientists and that this is one of the main payoffs of biorobotics. Indeed, technoscientists play an extremely important role in 21st-century culture and in the current critical production of knowledge. Today’s world is increasingly technological, or rather, it is a bio-hybrid system in which the biological and the technological are mixed. Therefore, studying the behaviour of robotic systems and the phenomena of animal-robot interaction means analysing, understanding, and shaping our world. Indeed, in the conclusion of the paper, we broadly reflect on the philosophical and disciplinary payoff of seeing biorobotics as a science and/or technoscience for the increasingly bio-hybrid and technical world of the 21st century.

Three-Dimensional Biohybrid StarPEG-Heparin Hydrogel Cultures for Modeling Human Neuronal Development and Alzheimer's Disease Pathology.

In this chapter, we present the methodology currently used in our laboratory to generate a starPEG-MMP (starPEG)- and heparin maleimide HM06 (heparin)-based 3D cell culture system, in a hydrogel, that can be used to study human neuronal development and Alzheimer's disease (AD) pathology. A 3D cell culture system can mimic the in vivo cellular environment better than a 2D format, in which these cells exhibit neural network formation, electrophysiological activity, tissue-specific extracellular matrix (ECM) deposition, and neurotransmitter responsiveness. When treated with amyloid beta-42 (Aβ42) peptides, this system recapitulates many of the pathological effects of AD, including reduced neural stem cell proliferation, impaired neuronal network formation, dystrophic axonal ends, synaptic loss, failure to deposit ECM, elevated tau hyperphosphorylation, and formation of neurofibrillary tangles. Culturing human primary cortical astrocyte (pHA)- or induced pluripotent stem cell (iPSC)-derived human neural stem cells in this biohybrid hydrogel system has led to the discovery of novel regulatory pathways underlying neurodegenerative pathology in different phases of AD.

Self-Assembled Nanoscale Manganese Oxides Enhance Carbon Capture by Diatoms.

Continuous CO2 emissions from human activities increase atmospheric CO2 concentrations and affect global climate change. The carbon storage capacity of the ocean is 20-fold higher than that of the land, and diatoms contribute to approximately 40% of carbon capture in the ocean. Manganese (Mn) is a major driver of marine phytoplankton growth and the marine carbon pump. Here, we discovered self-assembled manganese oxides (MnOx) for CO2 fixation in a diatom-based biohybrid system. MnOx shared key features (e.g., di-μ-oxo-bridged Mn-Mn) with the Mn4CaO5 cluster of the biological catalyst in photosystem II and promoted photosynthesis and carbon capture by diatoms/MnOx. The CO2 capture capacity of diatoms/MnOx was 1.5-fold higher than that of diatoms alone. Diatoms/MnOx easily allocated carbon into proteins and lipids instead of carbohydrates. Metabolomics showed that the contents of several metabolites (e.g., lysine and inositol) were positively associated with increased CO2 capture. Diatoms/MnOx upregulated six genes encoding photosynthesis core proteins and a key rate-limiting enzyme (Rubisco, ribulose 1,5-bisphosphate carboxylase-oxygenase) in the Calvin-Benson-Bassham carbon assimilation cycle, revealing the link between MnOx and photosynthesis. These findings provide a route for offsetting anthropogenic CO2 emissions and inspiration for self-assembled biohybrid systems for carbon capture by marine phytoplankton.

Efficient autonomous navigation for terrestrial insect-machine hybrid systems

While bio-inspired and biomimetic systems draw inspiration from living materials, biohybrid systems incorporate them with synthetic devices, allowing the exploitation of both organic and artificial advantages inside a single entity. In the challenging development of centimeter-scaled mobile robots serving unstructured territory navigations, biohybrid systems appear as a potential solution in the forms of terrestrial insect-machine hybrid systems, which are the fusion of living ambulatory insects and miniature electronic devices. Although their maneuver can be deliberately controlled via artificial electrical stimulation, these hybrid systems still inherit the insects’ outstanding locomotory skills, orchestrated by a sophisticated central nervous system and various sensory organs, favoring their maneuvers in complex terrains. However, efficient autonomous navigation of these hybrid systems is challenging. The struggle to optimize the stimulation parameters for individual insects limits the reliability and accuracy of navigation control. This study overcomes this problem by implementing a feedback control system with an insight into tunable navigation control for an insect-machine hybrid system based on a living darkling beetle. Via a thrust controller for acceleration and a proportional controller for turning, the system regulates the stimulation parameters based on the instantaneous status of the hybrid robot. While the system can provide an overall success rate of ∼ 71% for path-following navigations, fine tuning its control parameters could further improve the outcome’s reliability and precision to up to ∼ 94% success rate and ∼ 1/2 body length accuracy, respectively. Such tunable performance of the feedback control system provides flexibility to navigation applications of insect-machine hybrid systems.

Intracellular InP quantum dots facilitate the conversion of carbon dioxide to value-added chemicals in non-photosynthetic bacteria

The semiconductor material with low bio-toxicity and broad light absorption region is crucial for semiconductor-bacteria photosynthetic biohybrid systems. Here we introduce the low-toxic InP/ZnSe/ZnS quantum dots (QDs) into the non-photosynthetic bacteria to produce value-added chemicals. The QDs with ideal biocompatibility act as the light-harvesting semiconductor in the photosynthetic biohybrid systems. The results indicate that the QDs can enter the interior of bacteria and effectively promote the conversion of CO2 to acetate. The electrons generated intracellular present the primary contribution in the conversion process compared with the extracellular electrons transferred through the electron mediator. The total productivity of photosynthesis products can reach ∼ 0.89 mmol·L−1·h−1 with a quantum efficiency (QE) of 6–8%. Assisted with the genetically engineered E.coli, the obtained acetate is then converted into a high-value-added product. The hybrid photosynthetic system that integrated the QDs and non-photosynthetic bacteria exhibits great potential for large-scale solar-to-chemical conversion applications and environmental protection.

Live fish learn to anticipate the movement of a fish-like robot * *Contribution to Special Issue ‘Biohybrid Systems: Bioinspired and Biomimetic Robots Interacting with Living Organisms’ edited by Schmickl et al.

The ability of an individual to predict the outcome of the actions of others and to change their own behavior adaptively is called anticipation. There are many examples from mammalian species—including humans—that show anticipatory abilities in a social context, however, it is not clear to what extent fishes can anticipate the actions of their interaction partners or what the underlying mechanisms are for that anticipation. To answer these questions, we let live guppies (Poecilia reticulata) interact repeatedly with an open-loop (noninteractive) biomimetic robot that has previously been shown to be an accepted conspecific. The robot always performed the same zigzag trajectory in the experimental tank that ended in one of the corners, giving the live fish the opportunity to learn both the location of the final destination as well as the specific turning movement of the robot over three consecutive trials. The live fish’s reactions were categorized into a global anticipation, which we defined as relative time to reach the robot’s final corner, and a local anticipation which was the relative time and location of the live fish’s turns relative to robofish turns. As a proxy for global anticipation, we found that live fish in the last trial reached the robot’s destination corner significantly earlier than the robot. Overall, more than 50% of all fish arrived at the destination before the robot. This is more than a random walk model would predict and significantly more compared to all other equidistant, yet unvisited, corners. As a proxy for local anticipation, we found fish change their turning behavior in response to the robot over the course of the trials. Initially, the fish would turn after the robot, which was reversed in the end, as they began to turn slightly before the robot in the final trial. Our results indicate that live fish are able to anticipate predictably behaving social partners both in regard to final movement locations as well as movement dynamics. Given that fish have been found to exhibit consistent behavioral differences, anticipation in fish could have evolved as a mechanism to adapt to different social interaction partners.

The present and future of neural interfaces.

The 2020's decade will likely witness an unprecedented development and deployment of neurotechnologies for human rehabilitation, personalized use, and cognitive or other enhancement. New materials and algorithms are already enabling active brain monitoring and are allowing the development of biohybrid and neuromorphic systems that can adapt to the brain. Novel brain-computer interfaces (BCIs) have been proposed to tackle a variety of enhancement and therapeutic challenges, from improving decision-making to modulating mood disorders. While these BCIs have generally been developed in an open-loop modality to optimize their internal neural decoders, this decade will increasingly witness their validation in closed-loop systems that are able to continuously adapt to the user's mental states. Therefore, a proactive ethical approach is needed to ensure that these new technological developments go hand in hand with the development of a sound ethical framework. In this perspective article, we summarize recent developments in neural interfaces, ranging from neurohybrid synapses to closed-loop BCIs, and thereby identify the most promising macro-trends in BCI research, such as simulating vs. interfacing the brain, brain recording vs. brain stimulation, and hardware vs. software technology. Particular attention is devoted to central nervous system interfaces, especially those with application in healthcare and human enhancement. Finally, we critically assess the possible futures of neural interfacing and analyze the short- and long-term implications of such neurotechnologies.

Enhanced biomass growth of lactobacillus acidophilus with CdS-NPs under visible light irradiation: Semi-artificial biohybrid system

The bacterial biomass growth in a semi-artificial biohybrid system composed of cadmium sulfide nanoparticles (as a light-harvesting semiconductor) and Lactobacillus acidophilus (as the non-pathogenic electron acceptor bacterium) was investigated. The photo-induced electrons generated from cadmium sulfide nanoparticles within the culture medium efficiently enhanced the biomass growth (about 1.15-fold) and indicated a significant difference (P-value = 0.003 (<0.05)) after 48 h of incubation at 37 °C under white light-emitting diode lamp (12 W) compared to the dark control (i.e. culture grown without nanoparticles and light exposure). Scanning electron microscopy indicated the well-distributed spherical nanoparticles on the smooth surface of the bacterium. Energy-dispersive X-ray spectroscopy revealed the presence of nanoparticles on the bacterium cell surface. The findings of this study revealed that integrating cadmium sulfide nanoparticles with Lactobacillus acidophilus under visible light irradiation had a positive effect on bacterium growth.

Heparin-Peptide Nanogranules for Thrombosis-Actuated Anticoagulation.

Despite nearly a century of clinical use as a blood thinner, heparin's rapid serum clearance and potential to induce severe bleeding events continue to urge the development of more effective controlled delivery strategies. Subcutaneous depots that steadily release the anticoagulant into circulation represent a promising approach to reducing overdose frequency, sustaining therapeutic concentrations of heparin in plasma, and prolonging anticoagulant activity in a safe and effective manner. Subcutaneously deliverable heparin-peptide nanogranules that allow for long-lasting heparin bioavailability in the circulatory system, while enabling on-demand activation of heparin's anticoagulant effects in the thrombus microenvironment, are reported. Biophysical studies demonstrate this responsive behavior is due to the sequestration of heparin within self-assembling peptide nanofibrils and its mechanically actuated decoupling to elicit antithrombotic effects at the clotting site. In vivo studies show these unique properties converge to allow subcutaneous nanogranule depots to extend heparin serum concentrations for an order of magnitude longer than standard dosing regimens while enabling prolonged and controlled anticoagulant activity. This biohybrid delivery system demonstrates a potentially scalable platform for the development of safer, easier to administer, and more effective antithrombotic nanotechnologies.

On monocytes and lymphocytes biolens clustering by in flow holographic microscopy.

Live cells act as biological lenses and can be employed as real-world optical components in bio-hybrid systems. Imaging at nanoscale, optical tweezers, lithography and also photonic waveguiding are some of the already proven functionalities, boosted by the advantage that cells are fully biocompatible for intra-body applications. So far, various cell types have been studied for this purpose, such as red blood cells, bacterial cells, stem cells and yeast cells. White Blood Cells (WBCs) play a very important role in the regulation of the human body activities and are usually monitored for assessing its health. WBCs can be considered bio-lenses but, to the best of our knowledge, characterization of their optical properties have not been investigated yet. Here, we report for the first time an accurate study of two model classes of WBCs (i.e., monocytes and lymphocytes) by means of a digital holographic microscope coupled with a microfluidic system, assuming WBCs bio-lens characteristics. Thus, quantitative phase maps for many WBCs have been retrieved in flow-cytometry (FC) by achieving a significant statistical analysis to prove the enhancement in differentiation among sphere-like bio-lenses according to their sizes (i.e., diameter d) exploiting intensity parameters of the modulated light in proximity of the cell optical axis. We show that the measure of the low intensity area (S: ) in a fixed plane, is a feasible parameter for cell clustering, while achieving robustness against experimental misalignments and allowing to adjust the measurement sensitivity in post-processing. 2D scatterplots of the identified parameters (d-S) show better differentiation respect to the 1D case. The results show that the optical focusing properties of WBCs allow the clustering of the two populations by means of a mere morphological analysis, thus leading to the new concept of cell-optical-fingerprint avoiding fluorescent dyes. This perspective can open new routes in biomedical sciences, such as the chance to find optical-biomarkers at single cell level for label-free diagnosis.

Enzymatic and Microbial Electrochemistry: Approaches and Methods.

The coupling of enzymes and/or intact bacteria with electrodes has been vastly investigated due to the wide range of existing applications. These span from biomedical and biosensing to energy production purposes and bioelectrosynthesis, whether for theoretical research or pure applied industrial processes. Both enzymes and bacteria offer a potential biotechnological alternative to noble/rare metal-dependent catalytic processes. However, when developing these biohybrid electrochemical systems, it is of the utmost importance to investigate how the approaches utilized to couple biocatalysts and electrodes influence the resulting bioelectrocatalytic response. Accordingly, this tutorial review starts by recalling some basic principles and applications of bioelectrochemistry, presenting the electrode and/or biocatalyst modifications that facilitate the interaction between the biotic and abiotic components of bioelectrochemical systems. Focus is then directed toward the methods used to evaluate the effectiveness of enzyme/bacteria-electrode interaction and the insights that they provide. The basic concepts of electrochemical methods widely employed in enzymatic and microbial electrochemistry, such as amperometry and voltammetry, are initially presented to later focus on various complementary methods such as spectroelectrochemistry, fluorescence spectroscopy and microscopy, and surface analytical/characterization techniques such as quartz crystal microbalance and atomic force microscopy. The tutorial review is thus aimed at students and graduate students approaching the field of enzymatic and microbial electrochemistry, while also providing a critical and up-to-date reference for senior researchers working in the field.

Investigations on Cellular Uptake Mechanisms and Immunogenicity Profile of Novel Bio-Hybrid Nanovesicles.

In drug delivery, the development of nanovesicles that combine both synthetic and cellular components provides added biocompatibility and targeting specificity in comparison to conventional synthetic carriers such as liposomes. Produced through the fusion of U937 monocytes’ membranes and synthetic lipids, our nano-cell vesicle technology systems (nCVTs) showed promising results as targeted cancer treatment. However, no investigation has been conducted yet on the immunogenic profile and the uptake mechanisms of nCVTs. Hence, this study was aimed at exploring the potential cytotoxicity and immune cells’ activation by nCVTs, as well as the routes through which cells internalize these biohybrid systems. The endocytic pathways were selectively inhibited to establish if the presence of cellular components in nCVTs affected the internalization route in comparison to both liposomes (made up of synthetic lipids only) and nano-cellular membranes (made up of biological material only). As a result, nCVTs showed an 8-to-40-fold higher cellular internalization than liposomes within the first hour, mainly through receptor-mediated processes (i.e., clathrin- and caveolae-mediated endocytosis), and low immunostimulatory potential (as indicated by the level of IL-1α, IL-6, and TNF-α cytokines) both in vitro and in vivo. These data confirmed that nCVTs preserved surface cues from their parent U937 cells and can be rationally engineered to incorporate ligands that enhance the selective uptake and delivery toward target cells and tissues.

Light-Driven [FeFe] Hydrogenase Based H2 Production in E. coli: A Model Reaction for Exploring E. coli Based Semiartificial Photosynthetic Systems.

Biohybrid technologies like semiartificial photosynthesis are attracting increased attention, as they enable the combination of highly efficient synthetic light-harvesters with the self-healing and outstanding performance of biocatalysis. However, such systems are intrinsically complex, with multiple interacting components. Herein, we explore a whole-cell photocatalytic system for hydrogen (H2) gas production as a model system for semiartificial photosynthesis. The employed whole-cell photocatalytic system is based on Escherichia coli cells heterologously expressing a highly efficient, but oxygen-sensitive, [FeFe] hydrogenase. The system is driven by the organic photosensitizer eosin Y under broad-spectrum white light illumination. The direct involvement of the [FeFe] hydrogenase in the catalytic reaction is verified spectroscopically. We also observe that E. coli provides protection against O2 damage, underscoring the suitability of this host organism for oxygen-sensitive enzymes in the development of (photo) catalytic biohybrid systems. Moreover, the study shows how factorial experimental design combined with analysis of variance (ANOVA) can be employed to identify relevant variables, as well as their interconnectivity, on both overall catalytic performance and O2 tolerance.

3D Printed Biohybrid Microsystems

AbstractThis review is devoted to the role of 3D printing in the development of a new high‐tech field, smart biohybrid microsystems. The motivation behind the development of this field is the intention to integrate the capabilities of biological systems optimized in the course of evolution with the achievements of modern methods of forming micro‐ and nanostructures. Biohybrid technologies have demonstrated enormous potential for creating new materials and devices intended for a wide range of applications, primarily in the fields of medicine and biology. In this review, attention is focused on 3D printing as the main driver of the development of technologies for the formation of biohybrid systems. Methods featuring micro‐ and nanoresolution, which make it possible to efficiently integrate artificial structures and biosystems at the cellular level, are discussed. The formation of smart biohybrid microsystems—complex multifunctional devices designed for the medical treatment and study of tissues and organs in vitro and in vivo—is analyzed. Particular attention is given to the most promising, breakthrough trends in the development of biohybrid systems—microrobots, organ‐on‐a‐chip systems, and brain–computer interfaces. The main strategies in designing such systems and the near‐future opportunities offered by them are outlined.

Photobioelectrochemistry of intact photosynthetic bacteria: Advances and future outlook

Over the past decade, increasing research focus has been posed on the use of intact and viable photosynthetic microorganisms in biohybrid electrochemical systems. Various works have shown that these biotic/abiotic photoelectrochemical systems can be utilized to develop biophotovoltaics, biosensing devices, and to perform light-driven bioelectrosynthesis. This Emerging Opinions article highlights the accomplishments achieved in the last three years in the photobioelectrochemistry of intact photosynthetic bacteria. Specifically, the new approaches utilized to shed light on the mechanisms at the basis of microbial photobioelectrochemistry are discussed, together with fascinating solutions to artificially tune the photo-induced extracellular electron transfer. The most recent groundbreaking achievements toward the practical application of biohybrid photoelectrochemical systems are also highlighted, before presenting a future outlook for intact photosynthetic bacteria photobioelectrochemistry. • Multidisciplinary approaches shed light on photo-induced extracellular electron transfer. • Synthetic biology for engineered photosynthetic bacteria in bioelectrochemical systems. • Nanoparticles biosynthesis and redox polymers for facilitated electron transfer. • Intact photosynthetic bacteria for biosensing, biophotovoltaics, and bioelectrosynthesis.

Microliter Scale Synthesis of Luciferase-Encapsulated Polymersomes as Artificial Organelles for Optogenetic Modulation of Cardiomyocyte Beating.

Constructing artificial systems that effectively replace or supplement natural biological machinery within cells is one of the fundamental challenges underpinning bioengineering. At the sub‐cellular scale, artificial organelles (AOs) have significant potential as long‐acting biomedical implants, mimicking native organelles by conducting intracellularly compartmentalized enzymatic actions. The potency of these AOs can be heightened when judiciously combined with genetic engineering, producing highly tailorable biohybrid cellular systems. Here, the authors present a cost‐effective, microliter scale (10 µL) polymersome (PSome) synthesis based on polymerization‐induced self‐assembly for the in situ encapsulation of Gaussia luciferase (GLuc), as a model luminescent enzyme. These GLuc‐loaded PSomes present ideal features of AOs including enhanced enzymatic resistance to thermal, proteolytic, and intracellular stresses. To demonstrate their biomodulation potential, the intracellular luminescence of GLuc‐loaded PSomes is coupled to optogenetically engineered cardiomyocytes, allowing modulation of cardiac beating frequency through treatment with coelenterazine (CTZ) as the substrate for GLuc. The long‐term intracellular stability of the luminescent AOs allows this cardiostimulatory phenomenon to be reinitiated with fresh CTZ even after 7 days in culture. This synergistic combination of organelle‐mimicking synthetic materials with genetic engineering is therefore envisioned as a highly universal strategy for the generation of new biohybrid cellular systems displaying unique triggerable properties.