Bioreducible Amphiphilic Hyperbranched Polymer-Drug Conjugate for Intracellular Drug Delivery.

This paper reports synthesis of a bioreducible hyperbranched (HB) polymer by A2+B3 approach from commercially available dithiothreitol (DTT) (A2) and an easily accessible trifunctional monomer (B3) containing three reactive pyridyl-disulfide groups. Highly efficient thiol-activated disulfide exchange reaction leads to the formation of the HB polymer (Mw = 21000; D = 2.3) with bioreducible disulfide linkages in the backbone and two different functional groups, namely, hydroxyl and pyridyl-disulfide in the core and periphery, respectively, of the HB-polymer. Postpolymerization functionalization of the hydroxyl-groups with camptothecin (CPT), a topoisomerase inhibitor and known anticancer drug, followed by replacing the terminal pyridyl-disulfide groups with oligo-oxyethylene-thiol resulted in easy access to an amphiphilic HB polydisulfide-CPT conjugate (P1) with a very high drug loading content of ∼40%. P1 aggregated in water (above ∼10 µg/mL) producing drug-loaded nanoparticles (Dh ∼ 135 nm), which showed highly efficient glutathione (GSH)-triggered release of the active CPT. Mass spectrometry analysis of the GSH-treated P1 showed the presence of the active CPT drug as well as a cyclic monothiocarbonate product, which underpins the cascade-degradation mechanism involving GSH-triggered cleavage of the labile disulfide linkage, followed by intramolecular nucleophilic attack by the in situ generated thiol to the neighboring carbonate linkage, resulting in release of the active CPT drug. The P1 nanoparticle showed excellent cellular uptake as tested by confocal fluorescence microscopy in HeLa cells by predominantly endocytosis mechanism, resulting in highly efficient cell killing (IC50 ∼ 0.6 µg/mL) as evident from the results of the MTT assay, as well as the apoptosis assay. Comparative studies with an analogous linear polymer-CPT conjugate showed much superior intracellular drug delivery potency of the hyperbranched polymer.

Mitigation of Gas Porosity in Additive Manufacturing Using Experimental Data Analysis and Mechanistic Modeling.

Shielding gas, metal vapors, and gases trapped inside powders during atomization can result in gas porosity, which is known to degrade the fatigue strength and tensile properties of components made by laser powder bed fusion additive manufacturing. Post-processing and trial-and-error adjustment of processing conditions to reduce porosity are time-consuming and expensive. Here, we combined mechanistic modeling and experimental data analysis and proposed an easy-to-use, verifiable, dimensionless gas porosity index to mitigate pore formation. The results from the mechanistic model were rigorously tested against independent experimental data. It was found that the index can accurately predict the occurrence of porosity for commonly used alloys, including stainless steel 316, Ti-6Al-4V, Inconel 718, and AlSi10Mg, with an accuracy of 92%. In addition, experimental data showed that the amount of pores increased at a higher value of the index. Among the four alloys, AlSi10Mg was found to be the most susceptible to gas porosity, for which the value of the gas porosity index can be 5 to 10 times higher than those for the other alloys. Based on the results, a gas porosity map was constructed that can be used in practice for selecting appropriate sets of process variables to mitigate gas porosity without the need for empirical testing.