Eningeering bioinspired materials with controlled interactions with microorganisms

By re-engineering materials using bioinspired or “greener” chemistries, we can reduce or potentially eliminate toxic solvents, which would benefit the design of medical devices, water purification membranes, wearable electronics, as well as countless additional devices. In this presentation, I will discuss a story from each of my lab’s two synergistic research thrusts. First, I will discuss the effect that the fundamental properties of polymer coatings (i.e., molecular architecture, stiffness, and thickness) have on the adhesion of bacteria under quiescent conditions. By decoupling the effects of molecular architecture, stiffness, and thickness from coating chemistry, we have unlocked specific structure-property relationships that can be tailored to control the initial stage of bacterial adhesion. By understanding how materials properties influence bacterial adhesion, we may be able to decrease the concentration of commercial antibiotics needed to combat microbial biofilms. As an example of our work on green chemistry, I will highlight our recent research into nanofibers that form using only water and salt. Polyelectrolyte complexes (PECs) form due to the electrostatic complexation between oppositely-charged polymers. We have demonstrated that by exploiting the salt-driven plasticization of PECs, we can enable the electrospinning of robust fibers using an aqueous solution containing a pair of strong polyelectrolytes and salt. Electrospun PEC fibers are chemically and mechanically stable over a wide range of pH values, ionic strength conditions, and many organic solvents. The overall goal of this talk is to illustrate our recent findings and how these results can guide the green engineering of multifunctional materials

Encapsulating bacteria in alginate-based electrospun nanofibers

Encapsulation technologies are imperative for the safe delivery of live bacteria into the gut where they regulate bodily functions and human health. In this study, we develop alginate-based nanofibers that could potentially serve as a biocompatible, edible probiotic delivery system. By systematically exploring the ratio of three components, the biopolymer alginate (SA), the carrier polymer poly(ethylene oxide) (PEO), and the FDA approved surfactant polysorbate 80 (PS80), the surface tension and conductivity of the precursor solutions were optimized to electrospin bead-free fibers with an average diameter of 167 ± 23 nm. Next, the optimized precursor solution (2.8/1.2/3 wt% of SA/PEO/PS80) was loaded with Escherichia coli (E. coli, 108 CFU mL−1), which served as our model bacterium. We determined that the bacteria in the precursor solution remained viable after passing through a typical electric field (∼1 kV cm−1) employed during electrospinning. This is because the microbes are pulled into a sink-like flow, which encapsulates them into the polymer nanofibers. Upon electrospinning the E. coli-loaded solutions, beads that were much smaller than the size of an E. coli were initially observed. To compensate for the addition of bacteria, the SA/PEO/PS80 weight ratio was reoptimized to be 2.5/1.5/3. Smooth fibers with bulges around the live microbes were formed, as confirmed using fluorescence and scanning electron microscopy. By dissolving and plating the nanofibers, we found that 2.74 × 105 CFU g−1 of live E. coli cells were contained within the alginate-based fibers. This work demonstrates the use of electrospinning to encapsulate live bacteria in alginate-based nanofibers for the potential delivery of probiotics to the gut

Bacteria Adhesion Is Mechanosensitive To Polymer Gel Properties

Biofilm-related infections are a class of notoriously difficult to treat healthcare-associated infections, which commonly develop on the surface of implanted medical devices. Despite lacking visual, auditory, and olfactory perception, bacteria detect and settle on surfaces, however, how the intrinsic properties of materials affect the initial adhesion microorganisms remains relatively unknown. By unveiling the structure-property relationships between polymer materials and microbial adhesion, we could guide the design of materials a priori to resist the adhesion of infection causing microorganisms, such as Staphylococcus aureus. In this presentation, I will discuss the effect that fundamental properties of polymer coatings (i.e., molecular architecture, stiffness, and thickness) have on the surface-associated transport of bacteria and on the adhesion of bacteria under quiescent conditions. By decoupling the effects of molecular architecture, stiffness, and thickness from coating chemistry, we have unlocked specific structure-property relationships that can be tailored to control the degree of bacterial adhesion and subsequently, reduce the formation of biofilms. This work provides insight into the future design of bacteria-resistant surfaces with broad implications for applications including medical implants and equipment

Polyelectrolyte-Functionalized Nanofiber Mats Control the Collection and Inactivation of Escherichia coli

Quantifying the effect that nanofiber mat chemistry and hydrophilicity have on microorganism collection and inactivation is critical in biomedical applications. In this study, the collection and inactivation of Escherichia coli K12 was examined using cellulose nanofiber mats that were surface-functionalized using three polyelectrolytes: poly (acrylic acid) (PAA), chitosan (CS), and polydiallyldimethylammonium chloride (pDADMAC). The polyelectrolyte functionalized nanofiber mats retained the cylindrical morphology and average fiber diameter (~0.84 µm) of the underlying cellulose nanofibers. X-ray photoelectron spectroscopy (XPS) and contact angle measurements confirmed the presence of polycations or polyanions on the surface of the nanofiber mats. Both the control cellulose and pDADMAC-functionalized nanofiber mats exhibited a high collection of E. coli K12, which suggests that mat hydrophilicity may play a larger role than surface charge on cell collection. While the minimum concentration of polycations needed to inhibit E. coli K12 was 800 µg/mL for both CS and pDADMAC, once immobilized, pDADMAC-functionalized nanofiber mats exhibited a higher inactivation of E. coli K12, (~97%). Here, we demonstrate that the collection and inactivation of microorganisms by electrospun cellulose nanofiber mats can be tailored through a facile polyelectrolyte functionalization process

Linear Viscoelasticity and Time—Alcohol Superposition of Chitosan/Hyaluronic Acid Complex Coacervates

Complex coacervation is an associative liquid−liquid phase separation phenomenon resulting from the complexation of oppositely charged macroions. While it is well-known that the phase behavior and rheological character of the resulting coacervates can vary as a function of the identity of the various species present (i.e., macroions, salt, and solution conditions), the effect of solvent quality has been rarely studied. Here, the effect of adding small amounts of either methanol or ethanol to complex coacervates of the natural polymers chitosan and hyaluronic acid is described. The effect of cosolvent addition on the phase behavior and linear viscoelasticity of the resulting coacervates is characterized. Lastly, we explore the potential for using not only time−salt superposition but also time−alcohol and time−salt−alcohol superposition to provide insight into coacervate rheology

Understanding The Electrospinability Of Complex Coacervates

Complex coacervation is an associative, liquid-liquid phase separation that is driven by the electrostatic and entropic interactions between oppositely-charged polymers in water. For many coacervating systems it is possible to transition from the liquid coacervate state to a solid material by removing salt. This ‘saloplasticity’ allows for the processing of materials via methods such as spin coating, extrusion, etc. using the coacervate phase as a liquid precursor. In particular, we have developed an approach that uses complex coacervation as an environmentally friendly method for fabricating ultra-stable electrospun fibers directly from aqueous solutions. We have used this method to electrospin complexes of various synthetic polymers as well as natural biopolymers. These efforts have required the simultaneous exploration of the phase behavior of coacervate formation, as well as the rheology of the liquid coacervates

Electrospinning complex coacervates

As polymer-based materials become ever more integrated into our daily lives, there is an increasing need to develop both materials that are safe for the consumer, and manufacturing strategies that have a minimal impact on the environment. However, the vast majority of polymers require either organic solvents for dissolution, or the use of potentially cytotoxic cross-linking agents to prevent material dissolution. Additionally, many of the chemistries and solution conditions necessary for processing can damage cargo molecules and create biocompatibility issues for subsequent use. Complex coacervation is an associative, liquid-liquid phase separation that has the potential to circumvent many of the challenges associated with processing traditional polymers and encapsulating actives. Complex coacervation is driven by the electrostatic and entropic interactions between oppositely-charged polymers in water. For many coacervating systems, the solid or liquid nature of the complex can be tuned via the concentration of salt present. Additionally, the strength of the electrostatic interactions within the complex are such that in the absence of salt, solid complexes are highly resistant to thermal melting and/or solvent dissolution. Furthermore, complex coacervation has a strong history of use for the encapsulation of a range of cargo. We have taken advantage of this salt-driven plasticity to enable fabrication of ultra-stable electrospun fibers directly from aqueous solutions. These efforts have required the simultaneous characterization of coacervation, as well as the effect of cargo molecules on the phase behavior and rheology of the resulting coacervates/precursor solutions. Furthermore, these materials show tremendous promise for the use of electrospun coacervate-based nanofiber meshes across a range of applications

Current and Emerging Approaches to Engineer Antibacterial and Antifouling Electrospun Nanofibers

From ship hulls to bandages, biological fouling is a ubiquitous problem that impacts a wide range of industries and requires complex engineered solutions. Eliciting materials to have antibacterial or antifouling properties describes two main approaches to delay biofouling by killing or repelling bacteria, respectively. In this review article, we discuss how electrospun nanofiber mats are blank canvases that can be tailored to have controlled interactions with biologics, which would improve the design of intelligent conformal coatings or freestanding meshes that deliver targeted antimicrobials or cause bacteria to slip off surfaces. Firstly, we will briefly discuss the established and emerging technologies for addressing biofouling through antibacterial and antifouling surface engineering, and then highlight the recent advances in incorporating these strategies into electrospun nanofibers. These strategies highlight the potential for engineering electrospun nanofibers to solicit specific microbial responses for human health and environmental applications

Bacteria-Resistant, Transparent, Free-standing Films Prepared from Complex Coacervates

We report the fabrication, properties, and bacteria-resistance of polyelectrolyte complex (PEC) coatings and free-standing films. Poly(4-styrenesulfonic acid), poly(diallyldimethylammonium chloride), and salt were spin-coated into PEC films. After thermal annealing in a humid environment, highly transparent, mechanically strong, and chemically robust films were formed. Notably, we demonstrate that PEC coatings significantly reduce the attachment of Escherichia coli K12 without killing the microorganisms. We suggest that forming bacteria-resistant surface coatings from commercially available polymers holds the potential for use across a wide range of applications, including high-touch surfaces in medical settings

Mechanics of Intact Bone Marrow

The current knowledge of bone marrow mechanics is limited to its viscous properties, neglecting the elastic contribution of the extracellular matrix. To get a more complete view of the mechanics of marrow, we characterized intact yellow porcine bone marrow using three different, but complementary techniques: rheology, indentation, and cavitation. Our analysis shows that bone marrow is elastic, and has a large amount of intra- and inter-sample heterogeneity, with an effective Young’s modulus ranging from 0.25-24.7 kPa at physiological temperature. Each testing method was consistent across matched tissue samples, and each provided unique benefits depending on user needs. We recommend bulk rheology to capture the effects of temperature on tissue elasticity and moduli, indentation for quantifying local tissue heterogeneity, and cavitation rheology for mitigating destructive sample preparation. We anticipate the knowledge of bone marrow elastic properties for building in vitro models will elucidate mechanisms involved in disease progression and regenerative medicin