Browsing by Author "Fromen, Catherine A."
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Item Hydrogel nanoparticle degradation influences the activation and survival of primary macrophages(Journal of Materials Chemistry B, 2021-06-28) Jarai, Bader M.; Stillman, Zachary; Fromen, Catherine A.The effect of nanoparticle (NP) internalization on cell fate has emerged as an important consideration for nanomedicine design, as macrophages and other phagocytes are the primary clearance mechanisms of administered NP formulations. Pro-survival signaling is thought to be concurrent with phagocytosis and recent work has shown increased macrophage survival following lysosomal processing of internalized NPs. These observations have opened the door to explorations of NP physiochemical properties aimed at tuning the NP-driven macrophage survival at the lysosomal synapse. Here, we report that NP-induced macrophage survival and activation is strongly dependent on NP degradation rate using a series of thiol-containing poly(ethylene glycol) diacrylate-based NPs of equivalent size and zeta potential. Rapidly degrading, high thiol-containing NPs allowed for dramatic enhancement of cell longevity that was concurrent with macrophage stimulation after 2 weeks in ex vivo culture. While equivalent NP internalization resulted in suppressed caspase activity across the NP series, macrophage activation was correlated with increasing thiol content, leading to increased lysosomal activity and a robust pro-survival phenotype. Our results provide insight on tuning NP physiochemical properties as design handles for maximizing ex vivo macrophage longevity, which has implications for improving macrophage-based immune assays, biomanufacturing, and cell therapies.Item Macrophage variance: investigating how macrophage origin influences responses to soluble and physical cues with immortalized vs. primary cells in 2D and 3D culture(Frontiers in Biomaterials Science, 2024-05-22) Graf, Jodi; Bomb, Kartik; Trautmann-Rodriguez, Michael; Jarai, Bader M.; Gill, Nicole; Kloxin, April M.; Fromen, Catherine A.Macrophages are phagocytic innate immune cells capable of phenotypical switching in response to the local microenvironment. Studies often use either primary macrophages or immortalized cell lines for hypothesis testing, therapeutic assessment, and biomaterial evaluation without carefully considering the potential effects of cell source and tissue of origin, which strongly influence macrophage response. Surprisingly, limited information is available about how, under similar stimuli, immortalized cell lines and primary cells respond in both phenotypical and functional changes. To address this need, in this work, we cultured immortalized macrophage cell lines derived from different origins (i.e., blood, lung, peritoneal) to understand and compare macrophage phenotypical responses, including polarization and plasticity, morphological changes, and phagocytic functionalities, as well as compared primary macrophages extracted from peritoneal and bone marrow to their immortalized cell line counterparts. We found significant differences in baseline expression of different markers (e.g., CD86, MHCII, CD206, and EGR2) amongst different cell lines, which further influence both polarization and repolarization of the cells, in addition to their phagocytic functionality. Additionally, we observed that, while RAW 264.7 cells behave similarly to the primary bone marrow-derived macrophages, there are noticeable phenotypical and functional differences in cell line (IC-21) and primary peritoneal macrophages, highlighting tissue-specific differences in macrophage response amongst cell lines and primary cells. Moving to three-dimensional (3D) culture in well-defined biomaterials, blood-derived primary and cell line macrophages were encapsulated within hydrogel-based synthetic extracellular matrices and their polarization profiles and cell morphologies were compared. Macrophages exhibited less pronounced polarization during 3D culture in these compliant, soft materials compared to two-dimensional (2D) culture on rigid, tissue culture plastic plates. Overall, our findings highlight origin-specific differences in macrophage response, and therefore, careful considerations must be made to identify the appropriate cell source for the application of interest.Item Nanoparticle Internalization Promotes the Survival of Primary Macrophages(Wiley-VCH GmbH, 2022-02-09) Jarai, Bader M.; Fromen, Catherine A.; Bader M. Jarai, Catherine A. Fromen; Catherine A. Fromen; Bader M. JaraiMacrophages, a class of tissue resident innate immune cells, are responsible forsequestering foreign objects through the process of phagocytosis, making them apromising target for immune modulation via particulate engineering. Herein, it isreported that nanoparticle (NP) dosing and cellular internalization via phago-cytosis significantly enhance survival of ex vivo cultures of primary bone marrow-derived, alveolar, and peritoneal macrophages over particle-free controls. Theenhanced survival is attributed to suppression of caspase-dependent apoptosisand is linked to phagocytosis and lysosomal signaling. Uniquely, poly(ethyleneglycol)-based NP treatment extends cell viability in the absence of macrophagepolarization and enhances expression of prosurvival B cell lymphoma-2 (Bcl-2)protein in macrophages following multiple routes of in vivo administration. Theenhanced survival phenomenon is also applicable to NPs of alternative chem-istries, indicating the potential universality of this phenomenon with relevantdrug delivery particles. Thesefindings provide a framework for extending thelifespan of primary macrophages ex vivo for drug screening, vaccine studies,and cell therapies and have implications for in vivo particulate immune-engineering applications.Item Nanoparticle pre-treatment for enhancing the survival and activation of pulmonary macrophage transplant(Drug Delivery and Translational Research, 2023-03-14) Jarai, Bader M.; Bomb, Kartik; Fromen, Catherine A.Despite recent clinical successes of chimeric antigen receptor T cell therapies in treating liquid cancers, many lingering challenges stand in the way of therapeutic translation to broader types of malignancies. Macrophages have been proposed as alternatives to T cells given macrophages’ advantages in promoting tumor infiltration, acquiring diverse antigens, and possessing the ability to continuously stimulate adaptive responses. However, the poor survival of macrophages upon transplantation in addition to transient anti-tumor phenotypical states have been major obstacles standing in the way of macrophage-based cell therapies. Given recent discoveries of nanoparticle strategies in improving macrophage survival and promoting phenotype retention, we herein report the ability to extend the survival and phenotype of macrophage transplants in murine lungs via pre-treatment with nanoparticles of varying degradation rates. Macrophages pre-treated with 100 µg/ml dose of poly(ethylene glycol) diacrylate nanoparticle formulations improve pulmonary macrophage transplant survival over untreated cells beyond 7 days, where degradable nanoparticle formulations result in over a 50% increase in retention of transplanted cell counts relative to untreated cells. Furthermore, pre-treated macrophages more efficiently retain an imposed pro-inflammatory-like polarization state following transplantation out to 7 days compared to macrophages pre-treated with a classical pro-inflammatory stimulus, interferon-gamma, where CD86 costimulatory molecule expression is greater than 150% higher in pre-treated macrophage transplants compared to untreated counterparts. These findings provide an avenue for a major improvement in the lifespan and efficacy of macrophage-based cell therapies and have broader implications to other phagocyte-based cellular therapeutics and administration routes. Graphical abstract available at: https://doi.org/10.1007/s13346-023-01319-6Item Scalable 3D-printed lattices for pressure control in fluid applications(AIChE Journal, 2021-09-23) Woodward, Ian R.; Attia, Lucas; Patel, Premal; Fromen, Catherine A.Additive manufacturing affords precise control over geometries with high degrees of complexity and predefined structure. Lattices are one class of additive-only structures which have great potential in directing transport phenomena because they are highly ordered, scalable, and modular. However, a comprehensive description of how these structures scale and interact in heterogeneous systems is still undetermined. To advance this aim, we designed cubic and Kelvin lattices at two sub-5-mm length scales and compared published correlations to the experimental pressure gradient in pipes ranging from 12 to 52 mm diameter. We further investigated all combinations of the four lattices to evaluate segmented combinatorial behavior. The results suggest that a single correlation can describe pressure behavior for different lattice geometries and scales. Furthermore, combining lattice systems in series has a complex effect that is sensitive to part geometry. Together, these developments support the promise for tailored, modular lattice systems at laboratory scales and beyond.Item Scalable, process-oriented beam lattices: Generation, characterization, and compensation for open cellular structures(Additive Manufacturing, 2021-12-01) Woodward, Ian R.; Fromen, Catherine A.Additively manufactured lattices are emerging as promising candidates for structural, thermal, chemical, and biological applications. However, achieving a satisfactory prototype or final part with this level of complexity requires synthesis of disparate knowledge from the distinctly digital and physical processing stages. This work proposes an integrated framework for processing self-supporting, open lattice structures that do not require supports and facilitate material removal in post-processing steps. We describe a minimal yet comprehensive design strategy for generating uniform lattice structures with conformal open lattice skins for an arbitrary unit cell configuration. Using continuous liquid interface production (CLIP™) on a Carbon M1, printability is evaluated for five unique bending-dominated lattice structures at unit cell length scales from 0.5 to 3.5 mm and strut diameters ranging from 0.11 to 1.05 mm. Using a cubic lattice as a basis, we further examine dimensional fidelity with respect to 2D lattice void dimensions and part position, finding differences between length scales and within parts, due to physical processing artifacts. Finally, we demonstrate a functional grading strategy based on process control methods to compensate for dimensional deviations. Using an iterative approach based on a naïve process model, deviation of the planar strut radius in a cubic lattice was decreased by approximately 85% after two iterations. These insights and strategies can be readily applied to other structures, characterization techniques, and additive manufacturing processes, thereby improving the exchange of information between digital and physical processing and lowering the energy barriers to producing high-quality lattice parts.