Single-step 3D bioprinting of alginate-collagen I hydrogel fiber rings to promoter angiogenic network formation.

In the advent of tissue engineering and regenerative medicine, the demand for innovative approaches to biofabricate complex vascular structures is increasing. We describe a single-step 3D bioprinting method leveraging Aspect Biosystems RX1 technology, that integrates the crosslinking step at a flow-focusing junction, to biofabricate immortalized adult rat brain endothelial cell (SV-ARBEC)-encapsulated in alginate-collagen type I hydrogel rings, enabling robust angiogenesis and the formation of intricate vascular-like networks. This single-step biofabrication process involves the strategic layer-by-layer assembly of hydrogel rings, encapsulating SV-ARBECs in a spatially controlled manner while optimizing access to media and nutrients. The spatial arrangement of endothelial cells within the rings promotes angiogenic network formation and the organized development of vascular-like networks through facilitated constrained deposition of the cells within the hydrogel matrix forming tissue-like structures. This approach provides a platform that can be adapted to many different endothelial cell types and leveraged to better understanding the mechanisms driving angiogenesis and vascular-network formation in 3D bioprinted constructs supporting the development of more complex tissue and disease models for advancing drug discovery, tissue engineering and regenerative applications.

Characterization of Calcium- and Strontium-Polyphosphate Particles Toward Drug Delivery into Articular Cartilage.

Drug delivery into articular cartilage poses many challenges due in part to its lack of vasculature. While intra-articular injections are effective for the local administration of drugs, small molecules are rapidly cleared from the synovial fluid. As such, there is a need to develop effective drug delivery strategies to improve the residence times of bioactive molecules in the joint and elicit a sustained therapeutic effect. In this study, calcium- and strontium-polyphosphate particles are synthesized and characterized as potential drug carriers into articular cartilage. Physicochemical characterization reveals that the particles exhibit a spherical morphology, have a negative zeta potential, and are nanoscale in size. Biological characterization in chondrocytes confirms cellular uptake of the particles and demonstrates both size and concentration-dependent cytotoxicity at high concentrations. Furthermore, treatment of chondrocytes with these particles results in a reduction in cell proliferation and metabolic activity, confirming biological effects. Finally, incubation with cartilage tissue explants suggests successful uptake, despite the particles exhibiting a negative surface charge. Therefore, from the results of this study, these polyphosphate-based particles have potential as a drug carrier into articular cartilage and warrant further development.

Designing polymers for cartilage uptake: effects of architecture and molar mass.

Osteoarthritis (OA) is a progressive disease, involving the progressive breakdown of cartilage, as well as changes to the synovium and bone. There are currently no disease-modifying treatments available clinically. An increasing understanding of the disease pathophysiology is leading to new potential therapeutics, but improved approaches are needed to deliver these drugs, particularly to cartilage tissue, which is avascular and contains a dense matrix of collagens and negatively charged aggrecan proteoglycans. Cationic delivery vehicles have been shown to effectively penetrate cartilage, but these studies have thus far largely focused on proteins or nanoparticles, and the effects of macromolecular architectures have not yet been explored. Described here is the synthesis of a small library of polycations composed of N-(2-hydroxypropyl)methacrylamide (HPMA) and N-(3-aminopropyl)methacrylamide (APMA) with linear, 4-arm, or 8-arm structures and varying degrees of polymerization (DP) by reversible addition fragmentation chain-transfer (RAFT) polymerization. Uptake and retention of the polycations in bovine articular cartilage was assessed. While all polycations penetrated cartilage, uptake and retention generally increased with DP before decreasing for the highest DP. In addition, uptake and retention were higher for the linear polycations compared to the 4-arm and 8-arm polycations. In general, the polycations were well tolerated by bovine chondrocytes, but the highest DP polycations imparted greater cytotoxicity. Overall, this study reveals that linear polymer architectures may be more favorable for binding to the cartilage matrix and that the DP can be tuned to maximize uptake while minimizing cytotoxicity.

MicroRNA-124 Regulates Fatty Acid and Triglyceride Homeostasis.

MicroRNAs (miRNAs) are part of a complex regulatory network that modulates cellular lipid metabolism. Here, we identify miR-124 as a regulator of triglyceride (TG) metabolism. This study advances our knowledge of the role of miR-124 in human hepatoma cells. Transcriptional profiling of Huh7.5 cells overexpressing miR-124 reveals enrichment for host factors involved in fatty acid oxidation among repressed miRNA targets. In addition, miR-124 down-regulates arylacetamide deacetylase (AADAC) and adipose triglyceride lipase, lipases proposed to mediate breakdown of hepatic TG stores for lipoprotein assembly and mitochondrial ß-oxidation. Consistent with the inhibition of TG and fatty acid catabolism, miR-124 expression promotes cellular TG accumulation. Interestingly, miR-124 inhibits the production of hepatitis C virus, a virus that hijacks lipid pathways during its life cycle. Antiviral activity of miR-124 is consistent with repression of AADAC, a pro-viral host factor. Overall, our data highlight miR-124 as a novel regulator of TG metabolism in human hepatoma cells.