Glycopolymers are versatile materials for biomedical and healthcare applications, e.g., as carrier and polymeric therapeutics. Especially, the topology and surface composition of such materials play a key role in being promising materials in the anti-amyloidogenic interventions. Herein, 2nd and 3rd generation of pseudo-glycodendrimers (PGDs), based on hyperbranched polyester core with different sugar decorations, are synthesized, characterized, and used to investigate their anti-amyloidogenic properties toward Aβ (1-40) and (1-42), key players in Alzheimerś disease. The findings reveal that PGDs have a dendronized bottle brush architecture, as determined by SAXS analysis. PGDs are capable of interfering with the aggregation process of Amyloid-β peptides due to the high degree of sugar functionalization on the outer surface and the specific molecular shape. Additionally, cell viability studies indicate that PGDs exhibit concentration-dependent biocompatibility. Importantly, it is demonstrated that PGDs can be multi-functionalized by various sugar molecules, dyes, and/or peptides in a final one-pot approach. These findings suggest that PGDs may offer new avenues for therapeutic research in neurodegenerative diseases. Finally, it should be noted that this kind of highly branched glycopolymers possesses a molecular shape of dendronized bottle brushes and not a globular perfectly branched structure like glycodendrimers, as originally postulated.

Nanocatalytic medicine for treating cancer requires effective, versatile and novel tools and approaches to significantly improve the therapeutic efficiency for the interactions of (non-)enzymatic reactions. However, it is necessary to develop (non-)enzymatic nanotechnologies capable of selectively killing tumour cells without harming normal cells. Their therapeutic characteristics should be the adaption of tumours' extra- and intracellular environment to being specifically active. To contribute to this common goal, we propose the use of pH- and redox-responsive ferrocene containing polymersomes (FcPsomes). This allows the regulation of their biological activities for the controlled production of radicals via the Fenton reaction and the transport and release of cargo molecules via (destroying) host-guest interactions. This is provided by the Fc-membrane composition of FcPsomes showing different pH responsive active membrane, meaning the membrane permeability is triggering the Fenton reaction. The modulation of radical production is validated by various spectroscopic techniques. Moreover, these nanocontainers allow biological action of glucose oxidase (GOx) as therapeutic enzyme to produce glucono-1,5-lactone as proton source and hydrogen peroxide, initiating the Fenton reaction, in their interior. This provides a synergistic cancer therapeutic treatment for the starvation of hydrogen peroxide, but also ROS-mediated chemodynamic therapy at defined pH values. By modulating membrane permeability, we achieve environmentally regulated and locally driven therapeutic activity in the confinement of FcPsomes, ensuring specificity and safety treatments. The versatility of this platform extends beyond their specific application, allowing for their beneficial therapeutic use, for example, in signaling pathways of cells through the integration of various enzymes in FcPsomes.

Despite significant advances in cancer treatment, several challenges persist in optimizing effective cargo delivery, including enhancing bioavailability, improving targeted delivery, and overcoming biological barriers for improved tumor tissue penetration. There is an urgent need for versatile carriers capable of multi-functional targeting without compromising functionality. Here, we report a dual surface modification strategy to enhance the therapeutic efficacy of microrobotic platforms, through controlled, site-specific drug release. This dual functionalization integrates two distinct pH-sensitive polymeric nanoreservoirs with different membrane permeability. One nanoreservoir is engineered to release an antitumor agent -curcumin- in response to the acidic tumor microenvironment, while the second is designed to degrade the tumor extracellular matrix via enzymatic activity, facilitating enhanced diffussion of the therapeutic agent. This dual surface modification approach represents a significant advancement in the customizable integration of multifunctional nanoreservoirs. By leveraging dual compartmentalization, it prevents deactivation and cross-process interference, enabling precise nanoscale combination therapies for microrobotic cancer treatment. These surface-engineered microrobots hold promise for overcoming physiological barriers, ensuring stable cargo transport, and broadening the applicability of microrobotic platforms across diverse cancer types.

The membranization of membrane-less coacervates paves the way for the exploitation of complex protocells with regard to structural and cell-like functional behaviors. However, the controlled transformation from membranized coacervates to vesicles remains a challenge. This can provide stable (multi)phase and (multi)compartmental architectures through the reconfiguration of coacervate droplets in the presence of (bioactive) polymers, bio(macro)molecules and/or nanoobjects. Herein, we present a continuous protocell transformation from membrane-less coacervates to membranized coacervates and, ultimately, to giant hybrid vesicles. This transformation process is orchestrated by altering the balance of non-covalent interactions through varying concentrations of an anionic terpolymer, leading to dynamic processes such as spontaneous membranization of terpolymer nanoparticles at the coacervate surface, disassembly of the coacervate phase mediated by the excess anionic charge, and the redistribution of coacervate components in membrane. The diverse protocells during the transformation course provide distinct structural features and molecular permeability. Notably, the introduction of multiphase coacervates in this continuous transformation process signifies advancements toward the creation of synthetic cells with different diffusible compartments. Our findings emphasize the highly controlled continuous structural reorganization of coacervate protocells and represents a novel step toward the development of advanced and sophisticated synthetic protocells with more precise compositions and complex (membrane) structures.

In the rapidly advancing field of medical microrobotics, designing robots capable of addressing various challenges—such as imaging, biodegradation, and multifunctionality—is crucial. Departing from conventional research that often focuses on isolated aspects of microrobot functionality, this study presents an innovative approach to comprehensive microrobot design. Soft capsule microrobots that integrate capabilities such as magnetic navigation, autonomous maneuverability, in situ biodegradation, biosafe imaging, and drug delivery are reported. These microrobots are fabricated within the range of 20–120 μm, with a notable throughput of ≈102–103 microrobots per second. Furthermore, their locomotion performance has been demonstrated to remain stable for a period exceeding 10 h, all while employing real‐time optical closed‐loop control. The incorporation of ultrasound contrast agents not only amplifies imaging resolution but also ensures imaging contrast stability in a biological environment for over a period of 3 h. Second, the intentional integration of enzyme‐loaded nanometric polymersomes establishes a self‐contained, biodegradable system, accentuating the microrobots’ capacity to degrade without the addition of high enzyme concentrations. This integrated approach lays the groundwork for minimally invasive treatments toward personalized and targeted medicine.

The structure-function characteristics of isolated artificial organelles (AOs) in protocells are mainly known, but there are few reports on clustered or aggregated AOs. To imitate µm-sized complex and heterogeneous cell structures, approaches are needed that enable reversible changes in the aggregation state of colloidal structures in response to chemical, biological, and external stimuli. To construct adaptive organelle-like or cell-like reorganization characteristics, we present an advanced crosslinking strategy to fabricate clustered polymersomes as a platform based on host-guest interactions between azobenzene-containing polymersomes (Azo-Psomes) and a β-cyclodextrin-modified polymer (β-CD polymer) as a crosslinker. First, the reversible (dis)assembly of clustered Azo-Psomes is carried out by the alternating input of crosslinker and adamantane-PEG3000 as a decrosslinker. Moreover, cluster size dependence is demonstrated by environmental pH. These offer the controlled fabrication of various homogeneous and heterogeneous Azo-Psomes structures, including the size regulation and visualization of clustered AOs through a fluorescent enzymatic cascade reaction. Finally, a temperature-sensitive crosslinking agent with β-CD units can promote the coaggregation of Azo-Psomes mediated by temperature changes. Overall, these (co-)clustered Azo-Psomes and their successful transformation in AOs may provide new features for modelling biological systems for eukaryotic cells and systems biology.

Designing an electrochemical biosensor with the required features is a complex process involving multiple factors. For instance, nanocomposite materials used ensure the adaptation of the sensor to specific parameters on demand. But beyond making more versatile sensors, these materials likely offer excellent sensing platforms to mimic biological processes (e.g. cell communication) on the electrode surface. For this, developed methods are needed to integrate non-conductive nanoreactors for molecular communication into a biocompatible matrix on electrode surfaces with the request of spatially separated and controlled enzyme localization. Here, we present a novel reduced graphene oxide hybrid material to immobilize polymeric nanoreactors supported by an alginate network as a matrix on the electrode surface. The possibility of introducing cascade reactions in an electrochemical biosensor has the advantage of broadening the target substrates as well as their selectivity using enzymes in different nanocompartments (=compartmentalization). The polymeric vesicles allow the loading of enzymes or artificial enzymes, offering active center retention and long-term enzyme stability. Firstly, the inclusion of spatially separated polymeric active nanocompartments into the matrix on the electrode surface is used to monitor a simple enzyme reaction. However, the study’s accomplishment lies in its successful ability to monitor an enzyme cascade reaction, one producing and the other consuming hydrogen peroxide, hosting natural and artificial enzymes. This proof-of-concept generates an important contribution to the design of analytical platforms capable of detecting complex environments or even monitoring communication between cell-like structures, extremely useful for fields such as synthetic biology, biomimetics and advanced biosensors.

Objective: Increased mupirocin use leads to mupirocin resistance and is associated with persistence of methicillin-resistant Staphylococcus aureus (MRSA) carriers, prolonged hospitalization, and significant economic burdens for health systems. The study aimed to investigate the antimicrobial activity of compounds of Salvia rosmarinus L. ("rosemary", formerly Rosmarinus officinalis), alone or in combination with mupirocin, against multidrug-resistant MRSA using isolates obtained from pediatric patients. Methods: The in vitro antibacterial activity of the monoterpene alpha-pinene (alpha-Pi), a rosemary essential oil constituent, alone and in combination with mupirocin, was evaluated by determining the minimum inhibitory concentrations and minimum bactericidal concentrations (MBCs) and the fractional inhibitory concentration indices (FICIs) and fractional bactericidal concentration indices against multidrug-resistant clinical MRSA strains. The in vivo efficacy of alpha-Pi, alone and in combination with mupirocin, to eradicate MRSA infection was determined using an optimized mouse model of MRSA-infected wounds. Mouse skin samples (obtained via biopsy) were assessed for toxicity, and rabbit skin samples for irritation. Results: Both in vitro and in vivo, alpha-Pi was active against MRSA strains and acted synergistically with mupirocin against MRSA strains. Mupirocin-monoterpene combinations exhibited FICI values of 0.2 to 0.4, reducing the MBC of topical mupirocin 33-fold. A topical formulation containing alpha-Pi and mupirocin enhanced the efficacy of mupirocin in an in vivo MRSA-infected mouse skin model without significantly harming the skin of mice and rabbits. Conclusions: A topical formulation combining mupirocin and alpha-Pi may aid in the development of innovative agents for treating MRSA infections.

The design of compartments capable of carrying out biological reactions in a local space has provoked enormous interest by providing spatiotemporal and long-term selective control of biological activity. On the other hand, the application of metal-porphyrins in the field of biomedical science as nanozymes is gaining substantial importance. Porphyrins are the most widely studied tetrapyrrole-based compounds because of their important roles in vital biological processes and they possess peculiar photochemical, photophysical, and photo/redox properties. Herein, we demonstrate the use of pH-responsive and photo-crosslinked polymersomes for loading beta-cyclodextrin-Hemin complexes as potential peroxidase-mimicking cavity. The loading of catalytic active centers into polymeric vesicles represents a simple and effective strategy for enzyme mimicry. Physicochemical and enzyme-like properties are studied using a variety of characterization methods at different simulated microenvironments. This work offers an improvement of the aqueous solubility of the Hemin molecule, crucial for biomedical applications. In addition, these nanocompartments can be used as artificial radical-producing and hydrogen peroxide-consuming organelles, being able to replace cell functions in different microenvironments. Therefore, these artificial organelles, entrapping nanozymes, could provide promising synergistic and more personalized therapies on demand in modern nanomedicine.

The intricate nature of eukaryotic cells with intracellular compartments having differences in component concentration and viscosity in their lumen provides (membrane‐active) enzymes to trigger time‐ and concentration‐dependent processes in the intra‐/extracellular matrix. Herein, membrane‐active, enzyme‐loaded artificial organelles (AOs) are capitalized upon to develop fluidic and stable proteinaceous membrane‐based protocells. AOs in protocells induce the self‐assembly of oligopeptides into an artificial cytoskeleton that underlines their influence on the structure and functionality of protocells. A series of microscopical tools is used to validate the intracellular assembly and distribution of cytoskeleton, the changing protocells morphology, and AOs inclusion within cytoskeletal growth. Thus, the dynamics, diffusion, and viscosity of intracellular components in the presence of cytoskeleton are evaluated by fluorescence tools and enzymatic assay. Membrane‐active alkaline phosphatase in polymersomes as AOs fulfills the requirements of biomimetic eukaryotic cells to trigger intracellular environment, mobility, viscosity, diffusion, and enzymatic activity itself as well as high mechanical stability and high membrane fluidity of protocells. Thus membrane‐active AOs in protocells provide a variable reaction space in a changing intracellular environment and underline their regulatory role in the fabrication of complex protocell architectures and functions. This study contributes significantly to the effective biomimetics of cell‐like structures, shapes, and functions.

Various cellular functions are successfully mimicked, opening the door to the next generation of therapeutic approaches and systems biology. Herein, the first steps are taken toward the construction of artificial organelles for mimicking cell communication by docking and undocking of cargo in the membrane of swollen artificial organelles. Stimuli-responsive and crosslinked polymeric vesicles are used to allow docking processes at acidic pH at which ferrocene units in the swollen membrane state can undergo desired specific host-guest interaction using beta-cyclodextrin as model cargo. The release of the cargo mediated by two different enzymes, glucose oxidase and alpha-amylase, is investigated, triggered by distinct enzymatic undocking mechanisms. Different release times for a useful transport are shown that can be adapted to different communication pathways. In addition, Forster resonance energy transfer (FRET) experiments further support the hypotheses of host-guest inclusion complexation formation and their time-dependent breakdown. This work paves a way to a platform based on polymeric vesicles for synthetic biology, cell functions mimicking, and the construction of multifunctional cargo delivery system.

Responsive polymeric nanoparticles are an exciting field of study, yet little is known about the temperature sensitivity of those based on tertiary amine methacrylate block copolymers.

In recent years, there has been growing attention to designing synthetic protocells, capable of mimicking micrometric and multicompartmental structures and highly complex physicochemical and biological processes with spatiotemporal control. Controlling metabolism-like cascade reactions in coacervate protocells is still challenging since signal transduction has to be involved in sequential and parallelized actions mediated by a pH change. Herein, we report the hierarchical construction of membraneless and multicompartmentalized protocells composed of (i) a cytosol-like scaffold based on complex coacervate droplets stable under flow conditions, (ii) enzyme-active artificial organelles and a substrate nanoreservoir capable of triggering a cascade reaction between them in response to a pH increase, and (iii) a signal transduction component based on the urease enzyme capable of the conversion of an exogenous biological fuel (urea) into an endogenous signal (ammonia and pH increase). Overall, this strategy allows a synergistic communication between their components within the membraneless and multicompartment protocells and, thus, metabolism-like enzymatic cascade reactions. This signal communication is transmitted through a scaffold protocell from an "inactive state" (nonfluorescent protocell) to an "active state" (fluorescent protocell capable of consuming stored metabolites).

Post-loading of polymersomes with different bio(macro)molecules has been successfully demonstrated, thus mimicking the diffusion processes through biological membranes. However, it is still an open issue the extent to which this diffusion process leads to transmembrane transportation, or rather encapsulation of cargo within the membrane. In this study, well-established pH-responsive and cross-linked polymeric vesicles are studied. A pH-controllable and stable membrane, as well as a hollow particle shape and membrane uniformity are confirmed using dynamic light scattering (DLS) and cryogenic-transmission electron microscopy (Cryo-TEM). Post-loading with myoglobin (Mb) as a model enzyme is analyzed using multidetector asymmetrical flow field-flow fractionation (AF4). Advanced analysis of conformational parameters allowed for the estimation of enzyme localization and the pH-dependent loading efficiency thereof. Static light scattering coupled to AF4 is employed to successfully deliver information on the global size of the polymersomes (>50 nm). Furthermore, membrane structure and thickness, which are in the few nanometer range, can be successfully analyzed using small-angle neutron scattering (SANS). Deuterated solvent as well as Mb deuteration for tuning the contrast are considered. Two different vesicle model fits, as well as Kratky-Porod interpretation confirm effective determination of vesicle core and membrane thickness and for evaluation of the membrane changes after post-loading.

Structures and functions of eukaryotic cells with anouter permeablemembrane, a cytoskeleton, functional organelles, and motility canbe mimicked by giant multicompartment protocells containing varioussynthetic organelles. Herein, two kinds of artificial organelles withstimuli-triggered regulation ability, glucose oxidase-(GOx)-loadedpH-responsive polymersomes A (GOx-Psomes A) and urease-loaded pH-responsivepolymersomes B (Urease-Psomes B), and a pH-sensor (Dextran-FITC) areencapsulated into proteinosomes via the Pickering emulsion method.Thus, a polymersomes-in-proteinosome system is constructed which isable to probe biomimetic pH homeostasis. Alternating fuels (glucoseor urea) introduced from outside the protocell penetrate the membraneof proteinosomes and enter into GOx-Psomes A and Urease-Psomes B toproduce chemical signals (gluconic acid or ammonia) resulting in pH-feedbackloops (pH jump and pH drop). This will counteract the catalytic "switchon" or "switch off" of enzyme-loaded Psomes Aand B owing to their different pH-responsive membranes. Dextran-FITCin the proteinosome allows self-monitoring of slight pH fluctuationsin the lumen of protocells. Overall, this approach shows heterogeneouspolymersome-in-proteinosome architectures with sophisticated featuressuch as input-regulated pH changes mediated by negative and positivefeedback in loops and cytosolic pH self-monitoring, features thatare imperative for advanced protocell design.

Modern medical research develops interest in sophisticated artificial nano‐ and microdevices for future treatment of human diseases related to biological dysfunctions. This covers the design of protocells capable of mimicking the structure and functionality of eukaryotic cells. The authors use artificial organelles based on trypsin‐loaded pH‐sensitive polymeric vesicles to provide macrophage‐like digestive functions under physiological conditions. Herein, an artificial cell is established where digestive artificial organelles (nanosize) are integrated into a protocell (microsize). With this method, mimicking crossing of different biological barriers, capture of model protein pathogens, and compartmentalized digestive function are possible. This allows the integration of different components (e.g., dextran as stabilizing block) and the diffusion of pathogens in simulated cytosolic environment under physiological conditions. An integrated characterization approach is carried out, with identifying electrospray ionization mass spectrometry as an excellent detection method for the degradation of a small peptide such as β‐amyloid. The degradation of model enzymes is measured by enzyme activity assays. This work is an important contribution to effective biomimicry with the design of cell‐like functions having potential for therapeutic action.

Defects in cellular protein/enzyme encoding or even in organelles are responsible for many diseases. For instance, dysfunctional lysosome or macrophage activity results in the unwanted accumulation of biomolecules and pathogens implicated in autoimmune, neurodegenerative, and metabolic disorders. Enzyme replacement therapy (ERT) is a medical treatment that replaces an enzyme that is deficient or absent in the body but suffers from short lifetime of the enzymes. Here, this work proposes the fabrication of two different pH-responsive and crosslinked trypsin-loaded polymersomes as protecting enzyme carriers mimicking artificial organelles (AOs). They allow the enzymatic degradation of biomolecules to mimic simplified lysosomal function at acidic pH and macrophage functions at physiological pH. For optimal working of digesting AOs in different environments, pH and salt composition are considered the key parameters, since they define the permeability of the membrane of the polymersomes and the access of model pathogens to the loaded trypsin. Thus, this work demonstrates environmentally controlled biomolecule digestion by trypsin-loaded polymersomes also under simulated physiological fluids, allowing a prolonged therapeutic window due to protection of the enzyme in the AOs. This enables the application of AOs in the fields of biomimetic therapeutics, specifically in ERT for dysfunctional lysosomal diseases.

Anti-(ds)-DNA antibodies are the serological hallmark of Systemic Lupus Erythematosus (SLE). They assemble in the bloodstream with (ds)-DNA, forming immunocomplexes, which spread all over the body causing, among the other symptoms, lupic glomerulonephritis. Pathological manifestations of the disease may be reduced by destabilizing or inhibiting the formation of the immunocomplexes. In this respect, glycodendrimers showed peculiar interacting abilities towards this kind of biomolecule. Various generations of open-shell maltose-decorated poly(amidoamine) (PAMAM) and poly(propyleneimine) (PPI) dendrimers and two oligopeptides with different polyethylene glycol units were synthesized and characterized, and then tested for their anti-SLE activity. The activity of glycodendrimers and oligopeptides was evaluated in human plasma from patients with SLE, compared to healthy plasma, by means of an enzyme-linked immunosorbent assay (ELISA), and electron paramagnetic resonance (EPR) characterization using spin-label and spin-probe techniques. Different strategies for the immunocomplex formation were tested. The results show that both kinds of glycodendrimers and oligopeptides inhibited the formation of immunocomplexes. Also, a partial breakdown of preformed immunocomplexes was observed. Both ELISA and EPR analyses indicated a better activity of glycodendrimers compared to oligopeptides, the 3rd generation PPI dendrimer being the most promising against SLE. This study highlights the possibility to develop a new class of dendritic therapeutics for the treatment of Lupus in pre-clinical studies.

Robotic Microcarriers In article number 2204257, Oliver G. Schmidt, Mariana Medina-Sánchez, and co-workers report the development of 4D-printed multifunctional sperm-hybrid microcarriers containing soft and smart materials made with two-photon lithography. These multifunctional facets enable in situ sperm capacitation/hyperactivation, and the local degradation of the cumulus complex that surrounds the oocyte, both to facilitate the sperm–oocyte interaction for the ultimate goal of in vivo assisted fertilization.