Viscoelastic high-molecular-weight hyaluronic acid hydrogels support rapid glioblastoma cell invasion with leader-follower dynamics.

Hyaluronic acid (HA), the primary component of brain extracellular matrix, is increasingly used to model neuropathological processes, including glioblastoma (GBM) tumor invasion. While elastic hydrogels based on crosslinked low-molecular-weight (LMW) HA are widely exploited for this purpose and have proven valuable for discovery and screening, brain tissue is both viscoelastic and rich in high-MW (HMW) HA, and it remains unclear how these differences influence invasion. To address this question, hydrogels comprised of either HMW (1.5 MDa) or LMW (60 kDa) HA are introduced, characterized, and applied in GBM invasion studies. Unlike LMW HA hydrogels, HMW HA hydrogels relax stresses quickly, to a similar extent as brain tissue, and to a greater extent than many conventional HA-based scaffolds. GBM cells implanted within HMW HA hydrogels invade much more rapidly than in their LMW HA counterparts and exhibit distinct leader-follower dynamics. Leader cells adopt dendritic morphologies, similar to invasive GBM cells observed in vivo. Transcriptomic, pharmacologic, and imaging studies suggest that leader cells exploit hyaluronidase, an enzyme strongly enriched in human GBMs, to prime a path for followers. This study offers new insight into how HA viscoelastic properties drive invasion and argues for the use of highly stress-relaxing materials to model GBM.

A Balance between Inter- and Intra-Microgel Mechanics Governs Stem Cell Viability in Injectable Dynamic Granular Hydrogels.

Injectable hydrogels are increasingly explored for the delivery of cells to tissue. These materials exhibit both liquid-like properties, protecting cells from mechanical stress during injection, and solid-like properties, providing a stable 3D engraftment niche. Many strategies for modulating injectable hydrogels tune liquid- and solid-like material properties simultaneously, such that formulation changes designed to improve injectability can reduce stability at the delivery site. The ability to independently tune liquid- and solid-like properties would greatly facilitate formulation development. Here, such a strategy is presented in which cells are ensconced in the pores between microscopic granular hyaluronic acid (HA) hydrogels (microgels), where elasticity is tuned with static covalent intra-microgel crosslinks and flowability with mechanosensitive adamantane-cyclodextrin (AC) inter-microgel crosslinks. Using the same AC-free microgels as a 3D printing support bath, the location of each cell is preserved as it exits the needle, allowing identification of the mechanism driving mechanical trauma-induced cell death. The microgel AC concentration is varied to find the threshold from microgel yielding- to AC interaction-dominated injectability, and this threshold is exploited to fabricate a microgel with better injection-protecting performance. This delivery strategy, and the balance between intra- and inter-microgel properties it reveals, may facilitate the development of new cell injection formulations.

Lose the stress: Viscoelastic materials for cell engineering.

Biomaterials are widely used to study and control a variety of cell behaviors, including stem cell differentiation, organogenesis, and tumor invasion. While considerable attention has historically been paid to biomaterial elastic (storage) properties, it has recently become clear that viscous (loss) properties can also powerfully influence cell behavior. Here we review advances in viscoelastic materials for cell engineering. We begin by discussing collagen, an abundant naturally occurring biomaterial that derives its viscoelastic properties from its fibrillar architecture, which enables dissipation of applied stresses. We then turn to two other naturally occurring biomaterials that are more frequently modified for engineering applications, alginate and hyaluronic acid, whose viscoelastic properties may be tuned by modulating network composition and crosslinking. We also discuss the potential of exploiting engineered fibrous materials, particularly electrospun fiber-based materials, to control viscoelastic properties. Finally, we review mechanisms through which cells process viscous and viscoelastic cues as they move along and within these materials. The ability of viscoelastic materials to relax cell-imposed stresses can dramatically alter migration on two-dimensional surfaces and confinement-imposed barriers to engraftment and infiltration in three-dimensional scaffolds. STATEMENT OF SIGNIFICANCE: Most tissues and many biomaterials exhibit some viscous character, a property that is increasingly understood to influence cell behavior in profound ways. This review discusses the origin and significance of viscoelastic properties of common biomaterials, as well as how these cues are processed by cells to influence migration. A deeper understanding of the mechanisms of viscoelastic behavior in biomaterials and how cells interpret these inputs should aid the design and selection of biomaterials for specific applications.

Glioma Cells Secrete Collagen VI to Facilitate Invasion.

While glioblastoma (GBM) progression is associated with extensive extracellular matrix (ECM) secretion, the causal contributions of ECM secretion to invasion remain unclear. Here we investigate these contributions by combining engineered materials, proteomics, analysis of patient data, and a model of bevacizumab-resistant GBM. We find that GBM cells cultured in engineered 3D hyaluronic acid hydrogels secrete ECM prior to invasion, particularly in the absence of exogenous ECM ligands. Proteomic measurements reveal extensive secretion of collagen VI, and collagen VI-associated transcripts are correspondingly enriched in microvascular proliferation regions of human GBMs. We further show that bevacizumab-resistant GBM cells deposit more collagen VI than their responsive counterparts, which is associated with marked cell-ECM stiffening. COL6A3 deletion in GBM cells reduces invasion, ß-catenin signaling, and expression of mesenchymal markers, and these effects are amplified in hypoxia. Our studies strongly implicate GBM cell-derived collagen VI in microenvironmental remodeling to facilitate invasion.