RESUMO
Osteoarthritis (OA) poses significant therapeutic challenges, particularly OA that affects the hand. Currently available treatment strategies are often limited in terms of their efficacy in managing pain, regulating invasiveness, and restoring joint function. The APRICOTâ implant system developed by Aurora Medical Ltd (Chichester, UK) introduces a minimally invasive, bone-conserving approach for treating hand OA (https://apricot-project.eu/). By utilizing polycarbonate urethane (PCU), this implant incorporates a caterpillar track-inspired design to promote the restoration of natural movement to the joint. Surface modifications of PCU have been proposed for the biological fixation of the implant. This study investigated the biocompatibility of PCU alone or in combination with two surface modifications, namely dopamine-carboxymethylcellulose (dCMC) and calcium-phosphate (CaP) coatings. In a rat soft tissue model, native and CaP-coated PCU foils did not increase cellular migration or cytotoxicity at the implant-soft tissue interface after 3 d, showing gene expression of proinflammatory cytokines similar to that in non-implanted sham sites. However, dCMC induced an amplified initial inflammatory response that was characterized by increased chemotaxis and cytotoxicity, as well as pronounced gene activation of proinflammatory macrophages and neoangiogenesis. By 21 d, inflammation subsided in all the groups, allowing for implant encapsulation. In a rat bone model, 6 d and 28 d after release of the periosteum, all implant types were adapted to the bone surface with a surrounding fibrous capsule and no protracted inflammatory response was observed. These findings demonstrated the biocompatibility of native and CaP-coated PCU foils as components of APRICOTâ implants. STATEMENT OF SIGNIFICANCE: Hand osteoarthritis treatments require materials that minimize irritation of the delicate finger joints. Differing from existing treatments, the APRICOTâ implant leverages polycarbonate urethane (PCU) for minimally invasive joint replacement. This interdisciplinary, preclinical study investigated the biocompatibility of thin polycarbonate urethane (PCU) foils and their surface modifications with calcium-phosphate (CaP) or dopamine-carboxymethylcellulose (dCMC). Cellular and morphological analyses revealed that both native and Ca-P coated PCU elicit transient inflammation, similar to sham sites, and a thin fibrous encapsulation in soft tissues and on bone surfaces. However, dCMC surface modification amplified initial chemotaxis and cytotoxicity, with pronounced activation of proinflammatory and neoangiogenesis genes. Therefore, native and CaP-coated PCU possess sought-for biocompatible properties, crucial for patient safety and performance of APRICOTâ implant.
Assuntos
Fosfatos de Cálcio , Animais , Masculino , Ratos , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Dopamina/metabolismo , Dopamina/farmacologia , Materiais Revestidos Biocompatíveis/química , Materiais Revestidos Biocompatíveis/farmacologia , Cimento de Policarboxilato/química , Prótese Articular , Carboximetilcelulose Sódica/química , Carboximetilcelulose Sódica/farmacologia , Uretana/químicaRESUMO
Mucin glycoproteins are ideal biomacromolecules for drug delivery applications since they naturally offer a plethora of different functional groups that can engage in specific and unspecific binding interactions with cargo molecules. However, to fabricate drug carrier objects from mucins, suitable stabilization mechanisms have to be implemented into the nanoparticle preparation procedure that allow for drug release profiles that match the requirements of the selected cargo molecule and its particular mode of action. Here, we describe two different methods to prepare crosslinked mucin nanoparticles that can release their cargo either on-demand or in a sustained manner. This method chapter includes a description of the preparation and characterization of mucin nanoparticles (stabilized either with synthetic DNA strands or with covalent crosslinks generated by free radical polymerization), as well as protocols to quantify the release of a model drug from those nanoparticles.
Assuntos
Mucinas , Nanopartículas , Mucinas/metabolismo , Sistemas de Liberação de Medicamentos , Glicoproteínas , Portadores de Fármacos/química , Nanopartículas/químicaRESUMO
Whereas hydrogels created from synthetic polymers offer a high level of control over their stability and mechanical properties, their biomedical activity is typically limited. In contrast, biopolymers have evolved over billions of years to integrate a broad range of functionalities into a single design. Thus, biopolymeric hydrogels can show remarkable capabilities such as regulatory behavior, selective barrier properties, or antimicrobial effects. Still, despite their widespread use in numerous biomedical applications, achieving a meticulous control over the physical properties of macroscopic biopolymeric networks remains a challenge. Here, a macroscopic, DNA-crosslinked mucin hydrogel with tunable viscoelastic properties that responds to two types of triggers: temperature alterations and DNA displacement strands, is presented. As confirmed with bulk rheology and single particle tracking, the hybridized base pairs governing the stability of the hydrogel can be opened, thus allowing for a precise control over the hydrogel stiffness and even enabling a full gel-to-sol transition. As those DNA-crosslinked mucin hydrogels possess tunable mechanical properties and can be disintegrated on demand, they can not only be considered for controlled cargo release but may also serve as a role model for the development of smart biomedical materials in applications such as tissue engineering and wound healing.
Assuntos
Hidrogéis , Mucinas , Biopolímeros , Materiais Biocompatíveis , DNARESUMO
Macromolecular coatings can improve the surface properties of many medical devices by enhancing their wetting behavior, tribological performance, and anti-biofouling properties - and covalent coatings produced from mucin glycoproteins have been shown to be very powerful in all those aspects. However, obtaining highly functional mucin glycoproteins is, at the moment, still a time-consuming process, which renders mucins rather expensive compared to other biomacromolecules. Here, we study a set of commercially available macromolecules that have the potential of substituting mucins in coatings for endotracheal tubes (ETTs). We present an overview of the different properties these macromolecular coatings establish on the ETT surface and whether they withstand storage or sterilization processes. Our study pinpoints several strategies of how to enhance the lubricity of ETTs by applying macromolecular coatings but also demonstrates the limited anti-biofouling abilities of well-established macromolecules such as hyaluronic acid, polyethylene glycol, and dextran. Based on the obtained results, we discuss to what extent those coatings can be considered equivalent alternatives to mucin coatings for applications on medical devices - their applicability does not have to be limited to ETTs, but could be broadened to catheters and endoscopes as well.
Assuntos
Incrustação Biológica , Intubação Intratraqueal , Incrustação Biológica/prevenção & controle , Substâncias Macromoleculares , Mucinas , CatéteresRESUMO
With its potential to revolutionize the field of personalized medicine by producing customized medical devices and constructs for tissue engineering at low costs, 3D printing has emerged as a highly promising technology. Recent advancements have sparked increasing interest in the printing of biopolymeric hydrogels. However, owing to the limited printability of those soft materials, the lack of variability in available bio-inks remains a major challenge. In this study, a novel bio-ink is developed based on functionalized mucin-a glycoprotein that exhibits a multitude of biomedically interesting properties such as immunomodulating activity and strong anti-biofouling behavior. To achieve sufficient printability of the mucin-based ink, its rheological properties are tuned by incorporating Laponite XLG as a stabilizing agent. It is shown that cured objects generated from this novel bio-ink exhibit mechanical properties partially similar to that of soft tissue, show strong anti-biofouling properties, good biocompatibility, tunable cell adhesion, and immunomodulating behavior. The presented findings suggest that this 3D printable bio-ink has a great potential for a wide range of biomedical applications, including tissue engineering, wound healing, and soft robotics.
Assuntos
Bioimpressão , Tinta , Mucinas , Engenharia Tecidual , Impressão Tridimensional , Reologia , Hidrogéis/farmacologiaRESUMO
Biopolymer coatings on implants mediate the interactions between the synthetic material and its biological environment. Owing to its ease of preparation and the possibility to incorporate other bioactive molecules, layer-by-layer deposition is a method commonly used in the construction of biopolymer multilayers. However, this method typically requires at least two types of oppositely charged biopolymers, thus limiting the range of macromolecular options by excluding uncharged biopolymers. Here, we present a layer-by-layer approach that employs mussel-inspired polydopamine as the adhesive intermediate layer to build biopolymer multilayer coatings without requiring any additional chemical modifications. We select three biopolymers with different charge statesâanionic alginate, neutral dextran, and cationic polylysineâand successfully assemble them into mono-, double-, or triple-layers. Our results demonstrate that both the layer number and the polymer type modulate the coating properties. Overall, increasing the number of layers in the coatings leads to reduced cell attachment, lower friction, and higher drug loading capacity but does not alter the surface potential. Moreover, varying the biopolymer type affects the surface potential, macrophage differentiation, lubrication performance, and drug release behavior. This proof-of-concept study offers a straightforward and universal coating method, which may broaden the use of multilayer coatings in biomedical applications.
Assuntos
Quitosana , Dopamina , Liberação Controlada de Fármacos , Lubrificação , Quitosana/química , Biopolímeros/químicaRESUMO
Several challenges need to be overcome when applying nucleic acids as therapeutic agents. We developed a new way to control the onset of the release of cholesterol-conjugated oligonucleotides with a simple, versatile, and cheap platform. Moreover, we combine the platform into a dual-release system that can release a hydrophobic drug with zero-order kinetics, followed by a rapid release of cholesterol-conjugated DNA.
Assuntos
DNA , Oligonucleotídeos , Emulsões/química , DNA/química , ColesterolRESUMO
To achieve and maintain good operability of medical devices while reducing putative side effects for the patient, a promising strategy is to tailor the surface properties of such devices as they critically dictate the tissue compatibility and the biofouling behavior. Indeed, those properties can be strongly improved by generating mucin coatings on such medical devices. However, using coatings on optical systems, e.g., contact lenses, comes with various challenges: here, the geometrical and optical characteristics of the lens may not be compromised by either the coating process or the coating itself. In this study, we show how mucin macromolecules can be attached onto the surfaces of rigid, gas permeable contact lenses while maintaining all critical lens parameters. We demonstrate that the generated coatings improve the surface wettability (contact angles are reduced from 105° to 40° and liquid film break-up times are increased from <1 s to 31 s) and prevent tribological damage to corneal tissue. Additionally, such coatings are highly transparent (transmission values above 98 % compared to an uncoated sample are reached) and efficiently reduce lipid deposition to the lens surface by 90 % but fully maintain the geometrical and mechanical properties of the lenses. Thus, such mucin coatings could also be highly beneficial for other optical systems that are used in direct contact with tissues or body fluids.
Assuntos
Lentes de Contato , Mucinas , Propriedades de Superfície , Molhabilidade , Mucinas/química , Mucinas/farmacologia , Oxigênio , PermeabilidadeRESUMO
Syngas, a mixture of CO and H2 , is a high-priority intermediate for producing several commodity chemicals, e.g., ammonia, methanol, and synthetic hydrocarbon fuels. Accordingly, parallel sunlight-driven catalytic conversion of CO2 and protons to syngas is a key step toward a sustainable energy cycle. State-of-the-art catalytic systems and materials often fall short as application-oriented concurrent CO and H2 evolution requires challenging reaction conditions which can hamper stability, selectivity, and efficiency. Here a light-harvesting metal-organic framework hosting two molecular catalysts is engineered to yield colloidal, water-stable, versatile nanoreactors for photocatalytic syngas generation with highly controllable product ratios. In-depth fluorescence, X-ray, and microscopic studies paired with kinetic analysis show that the host delivers energy efficiently to active sites, conceptually yielding nanozymes. This unlocked sustained CO2 reduction and H2 evolution with benchmark turnover numbers and record incident photon conversions up to 36%, showcasing a highly active and durable all-in-one material toward application in solar energy-driven syngas generation.
RESUMO
Chemically fueled supramolecular materials offer unique properties that include spatial and temporal control and even the ability to self-heal. Indeed, a few studies have demonstrated the ability to self-heal, however, the underlying mechanisms remain unclear. Here, we designed a peptide that forms a fibrillar network upon chemical fueling. We were surprised that the hydrogel could self-heal despite the lack of dynamics in the fiber assembly and disassembly. We explain this behavior by a mechanism that involves the chemically fueled peptide molecules that cannot self-assemble due to the lack of nucleation sites. When the fibers are perturbed, new nucleation sites form that help the assembly resulting in the healing of the damaged network. Furthermore, we generalized the behavior for other peptides. We refer to this non-assembling, chemically-fueled peptide as a molecular glue. In future work, we aim to explore whether this self-healing mechanism applies to more complex structures, narrowing the gap between biological and synthetic self-assemblies.
RESUMO
Owing to the unhealthy lifestyle and genetic susceptibility of today's population, atherosclerosis is one of the global leading causes of life-threatening cardiovascular diseases. Although a rapid intervention is required for severe blood vessel constrictions, a systemic administration of anticoagulant drugs is not the preferred method of choice as the associated risk of bleeding complications is high. In this study, we present mechanosensitive nanogels that exhibit tunable degrees of disintegration upon exposure to different levels of stenosis. Those nanogels can be further functionalized to encapsulate charged drug molecules such as heparin, and they efficiently release their cargo when passing stenotic constrictions; however, passive drug leakage in the absence of mechanical shear stress is very low. Furthermore, heparin molecules liberated from those mechanosensitive nanogels show a similar blood clot lysis efficiency as the free drug molecules, which demonstrates that drug encapsulation into those nanogels does not interfere with the functionality of the cargo. Thus, the hemocompatible and mechanoresponsive nanogels developed here represent a smart and efficient drug delivery platform that can offer safer solutions for vascular therapy.
Assuntos
Sistemas de Liberação de Medicamentos , Heparina , Humanos , Nanogéis , Preparações Farmacêuticas , Constrição Patológica , Sistemas de Liberação de Medicamentos/métodos , Portadores de FármacosRESUMO
Mucus is a self-healing gel that lubricates the moist epithelium and provides protection against viruses by binding to viruses smaller than the gel's mesh size and removing them from the mucosal surface by active mucus turnover. As the primary nonaqueous components of mucus (≈0.2%-5%, wt/v), mucins are critical to this function because the dense arrangement of mucin glycans allows multivalence of binding. Following nature's example, bovine submaxillary mucins (BSMs) are assembled into "mucus-like" gels (5%, wt/v) by dynamic covalent crosslinking reactions. The gels exhibit transient liquefaction under high shear strain and immediate self-healing behavior. This study shows that these material properties are essential to provide lubricity. The gels efficiently reduce human immunodeficiency virus type 1 (HIV-1) and genital herpes virus type 2 (HSV-2) infectivity for various types of cells. In contrast, simple mucin solutions, which lack the structural makeup, inhibit HIV-1 significantly less and do not inhibit HSV-2. Mechanistically, the prophylaxis of HIV-1 infection by BSM gels is found to be that the gels trap HIV-1 by binding to the envelope glycoprotein gp120 and suppress cytokine production during viral exposure. Therefore, the authors believe the gels are promising for further development as personal lubricants that can limit viral transmission.
Assuntos
HIV-1 , Animais , Bovinos , Humanos , HIV-1/metabolismo , Herpesvirus Humano 2/metabolismo , Mucinas/metabolismo , Géis , Muco/metabolismoRESUMO
Biopolymer-based drug carriers are commonly used for the development of safe delivery systems. However, biopolymer-based systems are often highly sensitive to the acidic pH levels in the stomach and release most of their cargo before they have reached their point of destination. Such premature drug release combined with the resulting high dose requirements is not cost-efficient and comes with the risk of unwanted side effects on non-target tissues/organs. This problem can be mitigated by the mucin-based drug carriers developed here, which exhibit good stability at acidic pH levels as proven by dynamic light scattering and enzymatic degradation tests with pepsin. In addition, the mucin-based particles can deliver hydrophobic and hydrophilic drugs simultaneously, which is demonstrated both with experiments performed under in vitro sink conditions and with drug transport tests involving eukaryotic cells as targets. As photo-induced cross-links covalently stabilize those particles, they can release their payload over time in a sustained manner. The drug carrier system introduced here combines good stability with high drug encapsulation efficiency and very good biocompatibility and thus may be valuable for a broad spectrum of applications in biological settings.
Assuntos
Mucinas , Nanopartículas , Biopolímeros/metabolismo , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Mucinas/química , Nanopartículas/químicaRESUMO
The respiratory tract surface is protected from inhaled pathogens by a secreted layer of mucus rich in mucin glycoproteins. Abnormal mucus accumulation is a cardinal feature of chronic respiratory diseases, but the relationship between mucus and pathogens during exacerbations is poorly understood. We identified elevations in airway mucin 5AC (MUC5AC) and MUC5B concentrations during spontaneous and experimentally induced chronic obstructive pulmonary disease (COPD) exacerbations. MUC5AC was more sensitive to changes in expression during exacerbation and was therefore more predictably associated with viral load, inflammation, symptom severity, decrements in lung function, and secondary bacterial infections. MUC5AC was functionally related to inflammation, as Muc5ac-deficient (Muc5ac-/-) mice had attenuated RV-induced (RV-induced) airway inflammation, and exogenous MUC5AC glycoprotein administration augmented inflammatory responses and increased the release of extracellular adenosine triphosphate (ATP) in mice and human airway epithelial cell cultures. Hydrolysis of ATP suppressed MUC5AC augmentation of RV-induced inflammation in mice. Therapeutic suppression of mucin production using an EGFR antagonist ameliorated immunopathology in a mouse COPD exacerbation model. The coordinated virus induction of MUC5AC and MUC5B expression suggests that non-Th2 mechanisms trigger mucin hypersecretion during exacerbations. Our data identified a proinflammatory role for MUC5AC during viral infection and suggest that MUC5AC inhibition may ameliorate COPD exacerbations.
Assuntos
Mucina-5AC , Doença Pulmonar Obstrutiva Crônica , Trifosfato de Adenosina/metabolismo , Animais , Modelos Animais de Doenças , Humanos , Inflamação/metabolismo , Camundongos , Mucina-5AC/genética , Mucina-5AC/metabolismo , Mucina-5B/genética , Mucina-5B/metabolismo , Muco/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/virologia , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologiaRESUMO
With the "many-headed" slime mold Physarum polycelphalum having been voted the unicellular organism of the year 2021 by the German Society of Protozoology, we are reminded that a large part of nature's huge variety of life forms is easily overlooked - both by the general public and researchers alike. Indeed, whereas several animals such as mussels or spiders have already inspired many scientists to create novel materials with glue-like properties, there is much more to discover in the flora and fauna. Here, we provide an overview of naturally occurring slimy substances with adhesive properties and categorize them in terms of the main chemical motifs that convey their stickiness, i.e., carbohydrate-, protein-, and glycoprotein-based biological glues. Furthermore, we highlight selected recent developments in the area of material design and functionalization that aim at making use of such biological compounds for novel applications in medicine - either by conjugating adhesive motifs found in nature to biological or synthetic macromolecules or by synthetically creating (multi-)functional materials, which combine adhesive properties with additional, problem-specific (and sometimes tunable) features.
RESUMO
Conventional bioreactors are typically developed for the production of planktonic bacteria or submerged biofilms. In contrast, reactors for the continuous production of biofilms at the solid-air interface are scarce, and they require specific conditions since the bacteria need to attach firmly to the surface and require a permanent supply of moisture and nutrients from below. Recently, research from the field of civil engineering has pinpointed an increased need for the production of terrestrial biofilms: several variants of Bacillus subtilis biofilms have been shown to be useful additives to mortar that increase the water repellency, and, thus, the lifetime of the cementitious material. The bioreactor introduced here allows for the continuous production of such bacterial biofilms at the solid-air interface, and they have virtually identical properties as biofilms cultivated via classical microbiological techniques. This is made possible by equipping a rotating cylinder with a porous membrane that acts as a solid growth substrate the bacterial biomass can form on. In this configuration, nutrient supply is enabled via diffusive transport of a suitable growth medium from the core volume of the cylindrical reactor to the membrane surface. In addition to cultivating bacterial biofilms, the versatile and adaptable set up introduced here also enables the growth of other microbial organisms including the yeast Saccharomyces cerevisiae and the fungus Penicillium chrysogenum.
Assuntos
Biofilmes , Reatores Biológicos , Bactérias , Biomassa , Reatores Biológicos/microbiologia , FungosRESUMO
A fluent conversation with a virtual assistant, person-tailored news feeds, and deep-fake images created within seconds-all those things that have been unthinkable for a long time are now a part of our everyday lives. What these examples have in common is that they are realized by different means of machine learning (ML), a technology that has fundamentally changed many aspects of the modern world. The possibility to process enormous amount of data in multi-hierarchical, digital constructs has paved the way not only for creating intelligent systems but also for obtaining surprising new insight into many scientific problems. However, in the different areas of biosciences, which typically rely heavily on the collection of time-consuming experimental data, applying ML methods is a bit more challenging: Here, difficulties can arise from small datasets and the inherent, broad variability, and complexity associated with studying biological objects and phenomena. In this Review, we give an overview of commonly used ML algorithms (which are often referred to as "machines") and learning strategies as well as their applications in different bio-disciplines such as molecular biology, drug development, biophysics, and biomaterials science. We highlight how selected research questions from those fields were successfully translated into machine readable formats, discuss typical problems that can arise in this context, and provide an overview of how to resolve those encountered difficulties.
RESUMO
Drug delivery systems that release hydrophobic drugs with zero-order kinetics remain rare and are often complicated to use. In this work, we present a gellified emulsion (emulgel) that comprises oil droplets of a hydrolyzable oil entrapped in a hydrogel. In the oil, we incorporate various hydrophobic drugs and, because the oil hydrolyzes with zero-order kinetics, the release of the drugs is also linear. We tune the release period from three hours to 50 h by varying the initial oil concentration. We show that the release rate is tunable by varying the initial drug concentration. Our quantitative understanding of the system allows for predicting the drug release kinetics once the drug's partition coefficient between the oil and the aqueous phase is known. Finally, we show that our drug delivery system is fully functional after storing it at -20 °C. Cell viability studies show that the hydrolyzable oil and its hydrolysis product are non-toxic under the employed conditions. With its simplicity and versatility, our system is a promising platform for the zero-order release of the drug.
Assuntos
Óleos , Água , Liberação Controlada de Fármacos , Emulsões , Interações Hidrofóbicas e HidrofílicasRESUMO
Similar to how CRISPR has revolutionized the field of molecular biology, machine learning may drastically boost research in the area of materials science. Machine learning is a fast-evolving method that allows for analyzing big data and unveiling correlations that otherwise would remain undiscovered. It may hold invaluable potential to engineer novel functional materials with desired properties, a field, which is currently limited by time-consuming trial and error approaches and our limited understanding of how different material properties depend on each other. Here, we apply machine learning algorithms to classify complex biological materials based on their microtopography. With this approach, the surfaces of different variants of biofilms and plant leaves can not only be distinguished but also correctly classified according to their wettability. Furthermore, an importance ranking provided by one of the algorithms allows us to identify those surface features that are critical for a successful sample classification. Our study exemplifies how machine learning can contribute to the analysis and categorization of complex surfaces, a tool, which can be highly useful for other areas of materials science, such as damage assessment as well as adhesion or friction studies.
Assuntos
Algoritmos , Aprendizado de Máquina , Big Data , Biologia Molecular , Propriedades de SuperfícieRESUMO
Many supramolecular materials in biological systems are driven to a nonequilibrium state by the irreversible consumption of high-energy molecules such as ATP or GTP. As a result, they exhibit unique dynamic properties such as a tunable lifetime, adaptivity or the ability to self-heal. In contrast, synthetic counterparts that exist in or close to equilibrium are controlled by thermodynamic parameters and therefore lack these dynamic properties. To mimic biological materials more closely, synthetic self-assembling systems have been developed that are driven out of equilibrium by chemical reactions. This protocol describes the synthesis and characterization of such an assembly, which is driven by carbodiimide fuels. Depending on the amount of chemical fuel added to the material, its lifetime can be tuned. In the first step, the protocol details the synthesis and purification of the peptide-based precursors for the fuel-driven assemblies by solid-phase peptide synthesis. Then, we explain how to analyze the kinetic response of the precursors to a carbodiimide-based chemical fuel by HPLC and kinetic models. Finally, we detail how to study the emerging assembly's macro- and microscopic properties by time-lapse photography, UV-visible spectroscopy, shear rheology, confocal laser scanning microscopy and electron microscopy. The procedure is described using the example of a colloid-forming precursor Fmoc-E-OH and a fiber-forming precursor Fmoc-AAD-OH to emphasize the differences in characterization depending on the type of assembly. The characterization of a precursor's transient assembly can be done within 5 d. The synthesis and purification of a peptide precursor requires 2 d of work.