Implantable CAR T cell factories enhance solid tumor treatment.

Chimeric Antigen Receptor (CAR) T cell therapy has produced revolutionary success in hematological cancers such as leukemia and lymphoma. Nonetheless, its translation to solid tumors faces challenges due to manufacturing complexities, short-lived in vivo persistence, and transient therapeutic impact. We introduce 'Drydux' - an innovative macroporous biomaterial scaffold designed for rapid, efficient in-situ generation of tumor-specific CAR T cells. Drydux expedites CAR T cell preparation with a mere three-day turnaround from patient blood collection, presenting a cost-effective, streamlined alternative to conventional methodologies. Notably, Drydux-enabled CAR T cells provide prolonged in vivo release, functionality, and enhanced persistence exceeding 150 days, with cells transitioning to memory phenotypes. Unlike conventional CAR T cell therapy, which offered only temporary tumor control, equivalent Drydux cell doses induced lasting tumor remission in various animal tumor models, including systemic lymphoma, peritoneal ovarian cancer, metastatic lung cancer, and orthotopic pancreatic cancer. Drydux's approach holds promise in revolutionizing solid tumor CAR T cell therapy by delivering durable, rapid, and cost-effective treatments and broadening patient accessibility to this groundbreaking therapy.

Skeletal effects following developmental flame-retardant exposure are specific to sex and chemical class in the adult Wistar rat.

Introduction: Accumulating evidence reveals that endocrine disrupting chemicals (EDCs) can disrupt aspects of metabolic programming, suggesting that skeletal development may be at risk, a possibility that is rarely examined. The commercial flame retardant (FR) mixture, Firemaster 550 (FM 550), has repeatedly been shown to negatively influence metabolic programming, raising concerns that skeletal integrity may consequently be impaired. We have previously shown that gestational and lactational exposure to 1,000 µg FM 550 negatively affected sex-specific skeletal traits in male, but not female, rats assessed at 6 months of age. Whether this outcome is primarily driven by the brominated (BFR) or organophosphate ester (OPFR) portions of the mixture or the effects persist to older ages is unknown. Materials and methods: To address this, in the present study, dams were orally exposed throughout gestation and lactation to either 1,000 µg BFR, 1,000 µg OPFR, or 2,000 µg FM 550. Offspring (n = 8/sex/exposure) were weaned at PND 21 and assessed for femoral cortical and trabecular bone parameters at 8 months of age by high-resolution X-ray micro-computed tomography (micro-CT). Serum levels of serotonin, osteocalcin, alkaline phosphatase, and calcium were quantified. Results: FM 550 affected both sexes, but the females were more appreciably impacted by the OPFRs, while the males were more vulnerable to the BFRs. Conclusion: Although sex specificity was expected due to the sexual dimorphic nature of skeletal physiology, the mechanisms accounting for the male- and female-specific phenotypes remain to be determined. Future work aims to clarify these unresolved issues.

Low-dose intrapulmonary drug delivery device for studies on next-generation therapeutics in mice.

Although nebulizers have been developed for delivery of small molecules in human patients, no tunable device has been purpose-built for targeted delivery of modern large molecule and temperature-sensitive therapeutics to mice. Mice are used most of all species in biomedical research and have the highest number of induced models for human-relevant diseases and transgene models. Regulatory approval of large molecule therapeutics, including antibody therapies and modified RNA highlight the need for quantifiable dose delivery in mice to model human delivery, proof-of-concept studies, efficacy, and dose-response. To this end, we developed and characterized a tunable nebulization system composed of an ultrasonic transducer equipped with a mesh nebulizer fitted with a silicone restrictor plate modification to control the nebulization rate. We have identified the elements of design that influence the most critical factors to targeted delivery to the deep lungs of BALB/c mice. By comparing an in silico model of the mouse lung with experimental data, we were able to optimize and confirm the targeted delivery of over 99% of the initial volume to the deep portions of the mouse lung. The resulting nebulizer system provides targeted lung delivery efficiency far exceeding conventional nebulizers preventing waste of expensive biologics and large molecules during proof-of-concept and pre-clinical experiments involving mice. (Word Count =207).

Bioinstructive implantable scaffolds for rapid in vivo manufacture and release of CAR-T cells.

Despite their clinical success, chimeric antigen receptor (CAR)-T cell therapies for B cell malignancies are limited by lengthy, costly and labor-intensive ex vivo manufacturing procedures that might lead to cell products with heterogeneous composition. Here we describe an implantable Multifunctional Alginate Scaffold for T Cell Engineering and Release (MASTER) that streamlines in vivo CAR-T cell manufacturing and reduces processing time to a single day. When seeded with human peripheral blood mononuclear cells and CD19-encoding retroviral particles, MASTER provides the appropriate interface for viral vector-mediated gene transfer and, after subcutaneous implantation, mediates the release of functional CAR-T cells in mice. We further demonstrate that in vivo-generated CAR-T cells enter the bloodstream and control distal tumor growth in a mouse xenograft model of lymphoma, showing greater persistence than conventional CAR-T cells. MASTER promises to transform CAR-T cell therapy by fast-tracking manufacture and potentially reducing the complexity and resources needed for provision of this type of therapy.

Stretchable and Multi-Metal-Organic Framework Fabrics Via High-Yield Rapid Sorption-Vapor Synthesis and Their Application in Chemical Warfare Agent Hydrolysis.

Protocols to create metal-organic framework (MOF)/polymer composites for separation, chemical capture, and catalytic applications currently rely on relatively slow solution-based processing to form single MOF composites. Here, we report a rapid, high-yield sorption-vapor method for direct simultaneous growth of single and multiple MOF materials onto untreated flexible and stretchable polymer fibers and films. The synthesis utilizes favorable reactant absorption into polymers coupled with rapid vapor-driven MOF crystallization to form high surface area (>250 m2/gcomposite) composites, including UiO-66-NH2, HKUST-1, and MOF-525 on spandex, nylon, and other fabrics. The resulting composites are robust and maintain their functionality even after stretching. Stretchable MOF fabrics enable rapid solid-state hydrolysis of the highly toxic chemical warfare agent soman and paraoxon-methyl simulant. We show that this approach can readily be scaled by solution spray-coating of MOF precursors and to large area substrates.

Encapsulation of Electron Beam Melting Produced Alloy 718 to Reduce Surface Connected Defects by Hot Isostatic Pressing.

Defects in electron beam melting (EBM) manufactured Alloy 718 are inevitable to some extent, and are of concern as they can degrade mechanical properties of the material. Therefore, EBM-manufactured Alloy 718 is typically subjected to post-treatment to improve the properties of the as-built material. Although hot isostatic pressing (HIPing) is usually employed to close the defects, it is widely known that HIPing cannot close open-to-surface defects. Therefore, in this work, a hypothesis is formulated that if the surface of the EBM-manufactured specimen is suitably coated to encapsulate the EBM-manufactured specimen, then HIPing can be effective in healing such surface-connected defects. The EBM-manufactured Alloy 718 specimens were coated by high-velocity air fuel (HVAF) spraying using Alloy 718 powder prior to HIPing to evaluate the above approach. X-ray computed tomography (XCT) analysis of the defects in the same coated sample before and after HIPing showed that some of the defects connected to the EBM specimen surface were effectively encapsulated by the coating, as they were closed after HIPing. However, some of these surface-connected defects were retained. The reason for such remnant defects is attributed to the presence of interconnected pathways between the ambient and the original as-built surface of the EBM specimen, as the specimens were not coated on all sides. These pathways were also exaggerated by the high surface roughness of the EBM material and could have provided an additional path for argon infiltration, apart from the uncoated sides, thereby hindering complete densification of the specimen during HIPing.

Dual-energy computed tomography investigation of additive manufacturing aluminium-carbon-fibre composite joints.

In this work, aluminium-carbon-fibre reinforced plastic joints have been studied. Three types of samples were designed as double lap joints where the aluminium inserts were fabricated using both classical methods (milling) and additive manufacturing. Two versions of the joint were fabricated using additive manufacturing, one flat, and the other with small teeth designed to hook into the carbon-fibre plies. The joints were characterised using a non-linear, dual-energy computed tomography method to evaluate the bond between the composite and the metal inserts. The mechanical strength of the bonds was evaluated, both through tensile tests and four-point bending. A simple finite element model was used to discuss the joints behaviour. It was found that the joints fabricated using additive manufactured inserts were more resistant to peel stress than the milled inserts. In four-point bending tests the moment that the joint could withstand was increased by roughly 300% with the use of additive manufacturing and 400% with the use of additive manufacturing and small teeth. However, in tensile tests it was found that the teeth design reduced the maximum load capacity of the joints by roughly 30% due to porosity. Further, it was found that the additive manufactured samples did not add to the capability of withstanding shear stress. The information gained with the dual-energy computed tomography method was highly valuable as the behaviour of the joints would have been difficult to explain without the porosity information.

Additive Manufacturing of Polypropylene: A Screening Design of Experiment Using Laser-Based Powder Bed Fusion.

The use of commodity polymers such as polypropylene (PP) is key to open new market segments and applications for the additive manufacturing industry. Technologies such as powder-bed fusion (PBF) can process PP powder; however, much is still to learn concerning process parameters for reliable manufacturing. This study focusses in the process⁻property relationships of PP using laser-based PBF. The research presents an overview of the intrinsic and the extrinsic characteristic of a commercial PP powder as well as fabrication of tensile specimens with varying process parameters to characterize tensile, elongation at break, and porosity properties. The impact of key process parameters, such as power and scanning speed, are systematically modified in a controlled design of experiment. The results were compared to the existing body of knowledge; the outcome is to present a process window and optimal process parameters for industrial use of PP. The computer tomography data revealed a highly porous structure inside specimens ranging between 8.46% and 10.08%, with porosity concentrated in the interlayer planes in the build direction. The results of the design of experiment for this commercial material show a narrow window of 0.122 ≥ Ev ≥ 0.138 J/mm³ led to increased mechanical properties while maintaining geometrical stability.