Protocol for delivery of intraoperative immunotherapy to mice by surgical debulking of subcutaneous tumors.

Pre-clinical studies developing novel therapies to prevent cancer recurrence require appropriate surgical models. Here, we present a protocol for surgical debulking of subcutaneous tumors in mice, which allows for intraoperative application of immunotherapy-loaded biomaterials. We describe steps for inoculating tumor cells, anesthetizing mice, and performing surgery. We then detail procedures for applying biomaterial, bandaging mice, and data collection and analysis. The optimized bandaging regimen resolves the issue of wound dehiscence after surgery, for C57BL/6 mice, which interfere with surgical sites. For complete details on the use and execution of this protocol, please refer to Rwandamuriye et al.1.

Insulator-on-Conductor Fouling Amplifies Aqueous Electrolysis Rates.

The chemical industry is a major consumer of fossil fuels. Several chemical reactions of practical value proceed with the gain or loss of electrons, opening a path to integrate renewable electricity into chemical manufacturing. However, most organic molecules have low aqueous solubility, causing green and cheap electricity-driven reactions to suffer from intrinsically low reaction rates in industry's solvent of choice: water. Here, we show that a strategic, partial electrode fouling with hydrophobic insulators (oils and plastics) offsets kinetic limitations caused by poor reactant solubility, opening a new path for the direct integration of renewable electricity into the production of commodity chemicals. Through electrochemiluminescence microscopy, we reveal for the oxidation of organic reactants up to 6-fold reaction rate increase at the "fouled" oil-electrolyte-electrode interface relative to clean electrolyte-electrode areas. Analogously, electrodes partially masked (fouled) with plastic patterns, deposited either photolithographically (photoresists) or manually (inexpensive household glues and sealants), outperform clean electrodes. The effect is not limited to reactants of limited water solubility, and, for example, net gold electrodeposition rates are up to 22% larger at fouled than clean electrodes. In a system involving a surface-active reactant, rate augmentation is driven by the synergy between insulator-confined reactant enrichment and insulator-induced current crowding, whereas only the latter and possibly localized decrease in iR drop near the insulator are relevant in a system composed of non-surface-active species. Our counterintuitive electrode design enhances electrolysis rates despite the diminished area of intimate electrolyte-electrode contact and introduces a new path for upscaling aqueous electrochemical processes.

A surgically optimized intraoperative poly(I:C)-releasing hydrogel prevents cancer recurrence.

Recurrences frequently occur following surgical removal of primary tumors. In many cancers, adjuvant therapies have limited efficacy. Surgery provides access to the tumor microenvironment, creating an opportunity for local therapy, in particular immunotherapy, which can induce local and systemic anti-cancer effects. Here, we develop a surgically optimized biodegradable hyaluronic acid-based hydrogel for sustained intraoperative delivery of Toll-like receptor 3 agonist poly(I:C) and demonstrate that it significantly reduces tumor recurrence after surgery in multiple mouse models. Mechanistically, poly(I:C) induces a transient interferon alpha (IFNα) response, reshaping the tumor/wound microenvironment by attracting inflammatory monocytes and depleting regulatory T cells. We demonstrate that a pre-existing IFN signature predicts response to the poly(I:C) hydrogel, which sensitizes tumors to immune checkpoint therapy. The safety, immunogenicity, and surgical feasibility are confirmed in a veterinary trial in canine soft tissue tumors. The surgically optimized poly(I:C)-loaded hydrogel provides a safe and effective approach to prevent cancer recurrence.

Dose-Dependent Therapeutic Distinction between Active and Passive Targeting Revealed Using Transferrin-Coated PGMA Nanoparticles.

The paradigm of using nanoparticle-based formulations for drug delivery relies on their enhanced passive accumulation in the tumor interstitium. Nanoparticles with active targeting capabilities attempt to further enhance specific delivery of drugs to the tumors via interaction with overexpressed cellular receptors. Consequently, it is widely accepted that drug delivery using actively targeted nanoparticles maximizes the therapeutic benefit and minimizes the off-target effects. However, the process of nanoparticle mediated active targeting initially relies on their passive accumulation in tumors. In this article, it is demonstrated that these two tumor-targeted drug delivery mechanisms are interrelated and dosage dependent. It is reported that at lower doses, actively targeted nanoparticles have distinctly higher efficacy in tumor inhibition than their passively targeted counterparts. However, the enhanced permeability and retention effect of the tumor tissue becomes the dominant factor influencing the efficacy of both passively and actively targeted nanoparticles when they are administered at higher doses. Importantly, it is demonstrated that dosage is a pivotal parameter that needs to be taken into account in the assessment of nanoparticle mediated targeted drug delivery.