Microfluidic Assembly of Degradable, Stereocomplexed Hydrogel Microparticles.

Hydrogel microparticles (HMPs) have been investigated widely for their use in tissue engineering and drug delivery applications. However, translation of these highly tunable systems has been hindered by covalent cross-linking methods within microparticles. Stereocomplexation, a stereospecific form of physical cross-linking, provides a robust yet degradable alternative for creating translationally relevant HMPs. Herein, 4-arm polyethylene glycol (PEG) stars were used as macromolecular initiators from which oligomeric poly(l-lactic acid) (PLLA) was polymerized with a degree of polymerization (DPn) of 20 on each arm. Similarly, complementary propargyl-containing ABA cross-linkers with enantiomeric poly(d-lactic acid) (PDLA) segments (DPn = 20) on each arm. Droplets of these gel precursors were formed via a microfluidic organic-in-oil-in-water system where microparticles self-assembled via stereocomplexation and were stabilized after precipitation in deionized water. By varying the flow rate of the dispersed phase, well-defined microparticles with diameters of 33.7 ± 0.5, 62.4 ± 0.6, and 105.7 ± 0.8 µm were fabricated. Gelation due to stereocomplexation was confirmed via wide-angle X-ray scattering in which HMPs exhibited the signature diffraction pattern of stereocomplexed PLA at 2θ = 12.2, 21.2, 24.2°. Differential scanning calorimetry also confirmed stereocomplexation by the appearance of a crystallization exotherm (Tc = 37 °C) and a high-temperature endotherm (Tm = 159 °C) that does not appear in the homocrystallization of PLLA or the hydrogel precursors. Additionally, the propargyl handle present on the cross-linker allows for pre- or post-assembly thiol-yne "click" functionalization as demonstrated by the addition of thiol-containing fluorophores to the HMPs.

Factors affecting product association as a mechanism of host-cell protein persistence in bioprocessing.

Product association of host-cell proteins (HCPs) to monoclonal antibodies (mAbs) is widely regarded as a mechanism that can enable HCP persistence through multiple purification steps and even into the final drug substance. Discussion of this mechanism often implies that the existence or extent of persistence is directly related to the strength of binding but actual measurements of the binding affinity of such interactions remain sparse. Two separate avenues of investigation of HCP-mAb binding are reported here. One is the measurement of the affinity of binding of individual, commonly persistent Chinese hamster ovary (CHO) HCPs to each of a set of mAbs, and the other uses quantitative proteomic measurements to assess binding of HCPs in a null CHO harvested cell culture fluid (HCCF) to mAbs produced in the same cell line. The individual HCP measurements show that the binding affinities of individual HCPs to different mAbs can vary appreciably but are rarely very high, with only weak pH dependence. The measurements on the null HCCF allow estimation of individual HCP-mAb affinities; these are typically weaker than those seen in affinity measurements on isolated HCPs. Instead, the extent of binding appears correlated with the initial abundance of individual HCPs in the HCCF and the forms of the HCPs in the solution, i.e., whether HCPs are present as free molecules or as parts of large aggregates. Separate protein A chromatography experiments performed by feeding different fractions of a mAb-containing HCCF obtained by size-exclusion chromatography (SEC) showed clear differences in the number and identity of HCPs found in the protein A eluate. These results indicate a significant role for HCP-mAb association in determining HCP persistence through protein A chromatography, presumably through binding of HCP-mAb complexes to the resin. Overall, the results illustrate the importance of considering more fully the biophysical context of HCP-product association in assessing the factors that may affect the phenomenon and determine its implications. Knowledge of the abundances and the forms of individual or aggregated HCPs in HCCF are particularly significant, emphasizing the integration of upstream and downstream bioprocessing and the importance of understanding the collective properties of HCPs in addition to just the biophysical properties of individual HCPs.

Digital Light Processing to Afford High Resolution and Degradable CO2-Derived Copolymer Elastomers.

Vat photopolymerization 3D printing has proven very successful for the rapid additive manufacturing (AM) of polymeric parts at high resolution. However, the range of materials that can be printed and their resulting properties remains narrow. Herein, we report the successful AM of a series of poly(carbonate-b-ester-b-carbonate) elastomers, derived from carbon dioxide and bio-derived ϵ-decalactone. By employing a highly active and selective Co(II)Mg(II) polymerization catalyst, an ABA triblock copolymer (Mn=6.3 kg mol-1, ÐM=1.26) was synthesized, formulated into resins which were 3D printed using digital light processing (DLP) and a thiol-ene-based crosslinking system. A series of elastomeric and degradable thermosets were produced, with varying thiol cross-linker length and poly(ethylene glycol) content, to produce complex triply periodic geometries at high resolution. Thermomechanical characterization of the materials reveals printing-induced microphase separation and tunable hydrophilicity. These findings highlight how utilizing DLP can produce sustainable materials from low molar mass polyols quickly and at high resolution. The 3D printing of these functional materials may help to expedite the production of sustainable plastics and elastomers with potential to replace conventional petrochemical-based options.

Characterization and implications of host-cell protein aggregates in biopharmaceutical processing.

In the production of biopharmaceuticals such as monoclonal antibodies (mAbs) and vaccines, the residual amounts of host-cell proteins (HCPs) are among the critical quality attributes. In addition to overall HCP levels, individual HCPs may elude purification, potentially causing issues in product stability or patient safety. Such HCP persistence has been attributed mainly to biophysical interactions between individual HCPs and the product, resin media, or residual chromatin particles. Based on measurements on process streams from seven mAb processes, we have found that HCPs in aggregates, not necessarily chromatin-derived, may play a significant role in the persistence of many HCPs. Such aggregates may also hinder accurate detection of HCPs using existing proteomics methods. The findings also highlight that certain HCPs may be difficult to remove because of their functional complementarity to the product; specifically, chaperones and other proteins involved in the unfolded protein response (UPR) are disproportionately present in the aggregates. The methods and findings described here expand our understanding of the origins and potential behavior of HCPs in cell-based biopharmaceutical processes and may be instrumental in improving existing techniques for HCP detection and clearance.

Fabrication of Biomedical Scaffolds Using Biodegradable Polymers.

Degradable polymers are used widely in tissue engineering and regenerative medicine. Maturing capabilities in additive manufacturing coupled with advances in orthogonal chemical functionalization methodologies have enabled a rapid evolution of defect-specific form factors and strategies for designing and creating bioactive scaffolds. However, these defect-specific scaffolds, especially when utilizing degradable polymers as the base material, present processing challenges that are distinct and unique from other classes of materials. The goal of this review is to provide a guide for the fabrication of biodegradable polymer-based scaffolds that includes the complete pathway starting from selecting materials, choosing the correct fabrication method, and considering the requirements for tissue specific applications of the scaffold.

Sugar-Based Polymers with Stereochemistry-Dependent Degradability and Mechanical Properties.

Stereochemistry in polymers can be used as an effective tool to control the mechanical and physical properties of the resulting materials. Typically, though, in synthetic polymers, differences among polymer stereoisomers leads to incremental property variation, i.e., no changes to the baseline plastic or elastic behavior. Here we show that stereochemical differences in sugar-based monomers yield a family of nonsegmented, alternating polyurethanes that can be either strong amorphous thermoplastic elastomers with properties that exceed most cross-linked rubbers or robust, semicrystalline thermoplastics with properties comparable to commercial plastics. The stereochemical differences in the monomers direct distinct intra- and interchain supramolecular hydrogen-bonding interactions in the bulk materials to define their behavior. The chemical similarity among these isohexide-based polymers enables both statistical copolymerization and blending, which each afford independent control over degradability and mechanical properties. The modular molecular design of the polymers provides an opportunity to create a family of materials with divergent properties that possess inherently built degradability and outstanding mechanical performance.

Gradient versus End-Capped Degradable Polymer Sequence Variations Result in Stiff to Elastic Photochemically 3D-Printed Substrates.

Additive manufacturing affords the construction of complex scaffolds for tissue engineering, yet the limitation in material choice remains a barrier to clinical translation. Herein, a series of poly(propylene fumarate-co-propylene succinate) were synthesized using both one-pot and sequential ring-opening copolymerization reactions. Continuous liquid interface production-based photochemical 3D printing utilizing thiol-ene chemistry was used to fabricate precise structures with improved build time over the traditional poly(propylene fumarate)/diethyl fumarate 3D printing processes. Significantly, the materials do not exhibit a yield point under tension and Young's modulus of the 3D printed products can be tuned by more than 2 orders of magnitude (0.6-110 MPa) using polymer composition and the degree of polymerization. Printed constructs degrade fully under hydrolytic conditions and degradation rates can be tailored using polymer composition, polymer sequence, and resin formulation.

Shape Memory Behavior of Biocompatible Polyurethane Stereoelastomers Synthesized via Thiol-Yne Michael Addition.

Biodegradable shape memory elastomers have the potential for use in soft tissue engineering, drug delivery, and device fabrication applications. Unfortunately, few materials are able to meet the targeted degradation and mechanical properties needed for long-term implantable devices. In order to overcome these limitations, we have designed and synthesized a series of unsaturated polyurethanes that are elastic, degradable, and nontoxic to cells in vitro. The polymerization included a nucleophilic thiol-yne Michael addition between a urethane-based dipropiolate and a dithiol to yield an α,ß-unsaturated carbonyl moiety along the polymer backbone. The alkene stereochemistry of the materials was tuned between 32 and 82% cis content using a combination of an organic base and solvent polarity, which collectively direct the nucleophilic addition. The bulk properties such as tensile strength, modulus, and glass transition temperature can also be tuned broadly, and the hydrogen bonding imparted by the urethane moiety allows for these materials to elicit cyclic shape memory behavior. We also demonstrated that the in vitro degradation properties are highly dependent on the alkene stereochemistry.

Degradable, Photochemically Printable Poly(propylene fumarate)-Based ABA Triblock Elastomers.

Additive manufacturing is rapidly advancing tissue engineering, but the scope of its clinical translation is limited by a lack of materials designed to meet specific mechanical properties and resorption timelines. Materials that are printable via photochemical cross-linking, fully degradable, and elastomeric have proven to be particularly challenging to develop. Herein, we report the synthesis of a series of poly(propylene fumarate-b-γ-methyl-ε-caprolactone-b-propylene fumarate) ABA triblock polymers using sequential ring-opening polymerization and ring-opening copolymerization. When cross-linked photochemically using a continuous liquid interface production digital light processing Carbon M2 printer, these ABA-type triblock copolymers are durable elastomers with tunable degradation and elastic properties. The polymers are shown to undergo slow, hydrolytic degradation in vitro with minimal loss of mechanical performance during degradation.

Influence of Touch-Spun Nanofiber Diameter on Contact Guidance during Peripheral Nerve Repair.

Peripheral nerve regeneration across large gaps remains clinically challenging and scaffold design plays a key role in nerve tissue engineering. One strategy to encourage regeneration has utilized nanofibers or conduits to exploit contact guidance within the neural regenerative milieu. Herein, we report the effect of nanofiber topography on two key aspects of regeneration: Schwann cell migration and neurite extension. Substrates possessing distinct diameter distributions (300 ± 40 to 900 ± 70 nm) of highly aligned poly(ε-caprolactone) nanofibers were fabricated by touch-spinning. Cell migratory behavior and contact guidance were then evaluated both at the tissue level using dorsal root ganglion tissue explants and the cellular level using dissociated Schwann cells. Explant studies showed that Schwann cells emigrated significantly farther on fibers than control. However, both Schwann cells and neurites emigrated from the tissue explants directionally along the fibers regardless of their diameter, and the data were characterized by high variation. At the cellular level, dissociated Schwann cells demonstrated biased migration in the direction of fiber alignment and exhibited a significantly higher biased velocity (0.2790 ± 0.0959 µm·min-1) on 900 ± 70 nm fibers compared to other nanofiber groups and similar to the velocity found during explant emigration on 900 nm fibers. Therefore, aligned, nanofibrous scaffolds of larger diameters (900 ± 70 nm) may be promising materials to enhance various aspects of nerve regeneration via contact guidance alone. While cells track along with the fibers, this contact guidance is bidirectional along the fiber, moving in the plane of alignment. Therefore, the next critical step to direct regeneration is to uncover haptotactic cues that enhance directed migration.

Reassessing Undergraduate Polymer Chemistry Laboratory Experiments for Virtual Learning Environments.

Chemistry laboratory experiments are invaluable to students' acquisition of necessary synthetic, analytical, and instrumental skills during their undergraduate studies. However, the COVID-19 pandemic rendered face-to-face (f2f), in-person teaching laboratory experiences impossible from late 2019-2020 and forced educators to rapidly develop new solutions to deliver chemistry laboratory education remotely. Unfortunately, achieving learning and teaching objectives to the same caliber of in-person experiments is very difficult through distance learning. To overcome these hurdles, educators have generated many virtual and remote learning options for not only foundational chemistry courses but also laboratory experiments. Although the pandemic challenged high-level chemistry education, it has also created an opportunity for both students and educators to be more cognizant of virtual learning opportunities and their potential benefits within chemistry curriculum. Irrespective of COVID-19, virtual learning techniques, especially virtual lab experiments, can complement f2f laboratories and offer a cost-efficient, safe, and environmentally sustainable alternative to their in-person counterparts. Implementation of virtual and distance learning techniques-including kitchen chemistry and at-home laboratories, prerecorded videos, live-stream video conferencing, digital lab environment, virtual and augmented reality, and others-can provide a wide-ranging venue to teach chemistry laboratories effectively and encourage diversity and inclusivity in the field. Despite their relevance to real-world applications and potential to expand upon fundamental chemical principles, polymer lab experiments are underrepresented in the virtual platform. Polymer chemistry education can help prepare students for industrial and academic positions. The impacts of polymers in our daily life can also promote students' interests in science and scientific research. Hence, the translation of polymer lab experiments into virtual settings improves the accessibility of polymer chemistry education. Herein, we assess polymer experiments in the emergence of virtual learning environments and provide suggestions for further incorporation of effective polymer teaching and learning techniques into virtual settings.

Ultra-Tough Elastomers from Stereochemistry-Directed Hydrogen Bonding in Isosorbide-Based Polymers.

The remarkable elasticity and tensile strength found in natural elastomers are challenging to mimic. Synthetic elastomers typically feature covalently cross-linked networks (rubbers), but this hinders their reprocessability. Physical cross-linking via hydrogen bonding or ordered crystallite domains can afford reprocessable elastomers, but often at the cost of performance. Herein, we report the synthesis of ultra-tough, reprocessable elastomers based on linear alternating polymers. The incorporation of a rigid isohexide adjacent to urethane moieties affords elastomers with exceptional strain hardening, strain rate dependent behavior, and high optical clarity. Distinct differences were observed between isomannide and isosorbide-based elastomers where the latter displays superior tensile strength and strain recovery. These phenomena are attributed to the regiochemical irregularities in the polymers arising from their distinct stereochemistry and respective inter-chain hydrogen bonding.

Stereochemistry-Controlled Mechanical Properties and Degradation in 3D-Printable Photosets.

Stereochemistry provides an appealing handle by which to control the properties of small molecules and polymers. While it is established that stereochemistry in linear polymers affects their bulk mechanical properties, the application of this concept to photocurable networks could allow for resins that can accommodate the increasing demand for mechanically diverse materials without the need to significantly change their formulation. Herein, we exploit cis and trans stereochemistry in pre-resin oligomers to create photoset materials with mechanical properties and degradation rates that are controlled by their stereochemistry and molecular weight. Both the synthesis of stereopure (cis or trans) acrylate-terminated pre-polymers and the subsequent UV-triggered cross-linking occurred with a retention of stereochemistry, close to 100%. The stereochemistry of a 4 kDa oligomer within the resin enabled the tuning of the formulation to either a fast eroding, soft cis elastomer or a stiff trans plastic that is more resistant to degradation. These results demonstrate that stereochemistry is a powerful tool to modify the stiffness, toughness, and degradability of high-resolution, three-dimensional printed scaffolds from the same formulated ratio of components.

Polymeric Materials for Eye Surface and Intraocular Applications.

Ocular applications of polymeric materials have been widely investigated for medical diagnostics, treatment, and vision improvement. The human eye is a vital organ that connects us to the outside world so when the eye is injured, infected, or impaired, it needs immediate medical treatment to maintain clear vision and quality of life. Moreover, several essential parts of the eye lose their functions upon aging, causing diminished vision. Modern polymer science and polymeric materials offer various alternatives, such as corneal and scleral implants, artificial ocular lenses, and vitreous substitutes, to replace the damaged parts of the eye. In addition to the use of polymers for medical treatment, polymeric contact lenses can provide not only vision correction, but they can also be used as wearable electronics. In this Review, we highlight the evolution of polymeric materials for specific ocular applications such as intraocular lenses and current state-of-the-art polymeric systems with unique properties for contact lens, corneal, scleral, and vitreous body applications. We organize this Review paper by following the path of light as it travels through the eye. Starting from the outside of the eye (contact lenses), we move onto the eye's surface (cornea and sclera) and conclude with intraocular applications (intraocular lens and vitreous body) of mostly synthetic polymers and several biopolymers. Initially, we briefly describe the anatomy and physiology of the eye as a reminder of the eye parts and their functions. The rest of the Review provides an overview of recent advancements in next-generation contact lenses and contact lens sensors, corneal and scleral implants, solid and injectable intraocular lenses, and artificial vitreous body. Current limitations for future improvements are also briefly discussed.

Advancing Toward 3D Printing of Bioresorbable Shape Memory Polymer Stents.

Stents have evolved significantly since their introduction to the medical field in the early 1980s, becoming widely used in percutaneous coronary interventions and following nephrological procedures. However, the current commercially available stents do not degrade and remain in the body forever, leading to problems like restenosis in cardiovascular applications or requiring removal procedures in ureteral applications. Efforts to replace metal with resorbable materials have largely been halted after the commercial failure of and safety concerns elicited by Abbott's Absorb stent in 2017. Industry continues to use common polymers such as poly(l-lactide) (PLLA) and polycaprolactone (PCL) for biomedical products, but due to the weak mechanical properties of these bioresorbable materials in comparison to metals, these devices have struggled to accomplish the goals set, increasing risk of thrombosis. 3D printing stents using bioresorbable and shape memory materials could provide a method of patient-personalized production, remove the need for balloon expansion, and limit stent migration, thus bringing a new age of stent technology. The investigation of a range of 3D-printable and bioresorbable shape-memory polymers can provide solutions to the shortcomings of previously explored bioresorbable stents and revitalize the medical device industry efforts into advancing stent technology.

Zwitterion Surface-Functionalized Thermoplastic Polyurethane for Antifouling Catheter Applications.

Immobilizing zwitterionic molecules on material surfaces has been a promising strategy for creating antifouling surfaces. Herein, we show the ability to surface derivatize an allyl-ether-functionalized thermoplastic polyurethane (TPU) with a zwitterionic thiol in a radically induced thiol-ene reaction. The thermoplastic polyurethane was synthesized to have an allyl-ether side functionality using a modified chain extender molecule. The zwitterion surface functionalization was achieved via thiol-ene reaction in aqueous conditions. The presence of chemically tethered zwitterion moieties on the TPU surface was confirmed using X-ray photoelectron spectroscopy (XPS). Protein adsorption experiments via quartz crystal microbalance (QCM) show reduced fibrinogen attachment for the zwitterion-derivatized TPU when compared to its nonfunctionalized controls. The Zwitterion-TPU also showed a log scale reduction in bacterial adherence. For Pseudomonas aeruginosa and Staphylococcus epidermidis, the Zwitterion-TPU resulted in around a 40 and 50% lower bacterial biomass accumulation, respectively, over the time scale of the experiment. The fibroblast cell viability of TPU remained unaffected by functionalization with zwitterion thiol. The results from our model experiments suggest that a zwitterion-modified TPU is a promising candidate for antifouling catheters.

Amino Acid-Based Poly(ester urea)s as a Matrix for Extended Release of Entecavir.

The use of polymers as excipients for drug delivery has afforded stable formulations that reliably control the release of active pharmaceutical ingredients (APIs). While many materials are available and used, few polymers exhibit the numerous advantages, including amorphous characteristics, noninflammatory properties, and resorbable degradation products, like those of poly(ester urea)s (PEUs). Furthermore, stability issues that arise in various APIs can make formulation optimization difficult. Herein, a series of PEUs were synthesized that vary by the fraction of l-phenylalanine monomer incorporated into the copolymerization. The various PEUs and entecavir monohydrate were dry-mixed at different weight percentages (15, 30, and 50%). Filaments of the PEU formulations were extruded and analyzed quantitatively for drug loading and content uniformity by using µ-CT and UPLC analysis. Drug dissolution profiles from filament segments were monitored over a 4-week period and ultimately showed that the controlled release of entecavir was influenced by the incorporation of the l-phenylalanine within the polymer.

Tuning Cooperative Assembly with Bottlebrush Block Co-polymers for Porous Metal Oxide Films Using Solvent Mixtures.

Block copolymer templating enables the generation of well-defined pore sizes and geometries in a wide variety of frameworks, typically through evaporation-induced self-assembly (EISA). Here, we systematically modulate the solvent quality with mixtures of tetrahydrofuran-ethanol (THF-EtOH) to manipulate the unimer/micelle ratio in the precursor solution to explore how the associated solution structure influences the final pore morphology. A bottlebrush block copolymer (BBCP) with poly(ethylene oxide) and poly(t-butyl acrylate) side chains was used as the template for pore formation. Irrespective of the solvent composition, a bimodal pore size distribution was obtained with mesopores templated by small aggregates of the BBCP unimers (potentially low aggregation number micelles) and macropores templated by large self-assembled BBCP micelles. The morphology and pore characteristics of the metal oxide films were dependent on the THF-EtOH composition. Interestingly, an intermediate solvent composition where the volume of micelles is approximately half the volume of unimers (in the precursor solution) leads to the best ordering of micelle-templated pores and also the maximum porosity in the films. The micelle/unimer ratios in the precursor solutions do not correspond directly to the bimodal pore distribution in the metal oxide films, which we attribute to kinetically trapped assembly of the BBCP at a low THF content. The increased critical micelle concentration at high THF composition leads to changes in the unimer/micelle ratio during solvent evaporation. These results appear to be universal for a number of metal oxides (cobalt, magnesium, and zinc) with the porosity maximized at a THF/EtOH ratio of 3:1. These results suggest the potential for enhancements in the porosity of block copolymer-templated films by EISA methods through judicious solvent selection.

Postfabrication Tethering of Molecular Gradients on Aligned Nanofibers of Functional Poly(ε-caprolactone)s.

Substrates with combinations of topographical and biochemical cues are highly useful for a number of fundamental biological investigations. Tethered molecular concentration gradients in particular are highly desired for a number of biomedical applications including cell migration. Herein, we report a versatile method for the fabrication of aligned nanofiber substrates with a tunable concentration gradient along the fiber direction. 4-Dibenzocyclooctynol (DIBO) was used as an initiator for the ring-opening copolymerization of ε-caprolactone (εCL) and allyl-functionalized ε-caprolactone (AεPCL), which yielded a well-defined polymer with orthogonal functional handles. These materials were fabricated into aligned nanofiber substrates via touch-spinning. Fibers were modified post-spinning with a concentration gradient of fluorescently labeled dye via a light activated thiol-ene reaction through a photomask. As a demonstration, the cell adhesive peptide RGD was chemically tethered to the fiber surface at a second functionalization site via strain-promoted azide-alkyne cycloaddition (SPAAC). This novel approach affords fabrication of dual functional nanofiber substrates.

3D Printing of Poly(propylene fumarate) Oligomers: Evaluation of Resin Viscosity, Printing Characteristics and Mechanical Properties.

Complex three-dimensional (3D) pore geometries, useful for tissue engineering scaffolds, can be fabricated via photo-crosslinking of resorbable poly(propylene fumarate) (PPF) resins using stereolithography (SLA) and/or continuous digital light processing (cDLP) methods. Physico-chemical parameters inherent to 3D printable resin design, include viscosity, polymer concentration, degree of polymerization, and resin printing temperature. We report here on our study of these parameters and their influence the cDLP 3D printing process and the resulting mechanical properties. A series of PPF oligomers were synthesized by the ring-opening copolymerization (ROCOP) of maleic anhydride and propylene oxide followed by a base-catalyzed isomerization. The resin viscosities were measured as a function of number-average molecular mass ([Formula: see text]) of the PPF oligomers (1.1, 1.7 and 2.0 kDa), concentrations of PPF in the reactive diluent diethyl fumarate (DEF) (50 and 75 wt %) and resin temperature (25 to 55 °C). The zero-shear viscosity (η0) of the resins was found to be temperature-dependent and follow a linear Arrhenius relationship. Tensile tests demonstrated mechanical properties within the range of trabecular bone, with the ultimate strength at break above 15 MPa and elastic moduli between 178 and 199 MPa.