3-Dimensional spatially organized PEG-based hydrogels for an aortic valve co-culture model

Abstract

Physiologically relevant in vitro models are needed to study disease progression and to develop and screen potential therapeutic interventions for disease. Heart valve disease, in particular, has no early intervention or non-invasive treatment because there is a lack of understanding the cellular mechanisms which lead to disease. Here, we establish a novel, customizable synthetic hydrogel platform that can be used to study cell-cell interactions and the factors which contribute to valve disease. Spatially localized cell adhesive ligands bound in the scaffold promote cell growth and organization of valve interstitial cells and valve endothelial cells in 3D co-culture. Both cell types maintained phenotypes, homeostatic functions, and produced zonally localized extracellular matrix. This model extends the capabilities of in vitro research by providing a platform to perform direct contact co-culture with cells in their physiologically relevant spatial arrangement.

Myofibroblastic activation of valvular interstitial cells is modulated by spatial variations in matrix elasticity and its organization

“…Amongst these factors, elastic modulus has been widely studied in marrow-derived mesenchymal stem cells and shown to influence cell-matrix interactions, morphology and cell fate via mechanotransduction [54,55,56]. Previous studies show that cells sense the ECM through focal adhesions and cytoskeleton tension, and furthermore, matrix mechanics direct cell fate [57,58].…”

Hyaluronan Hydrogels for a Biomimetic Spongiosa Layer of Tissue Engineered Heart Valve Scaffolds

“…22,25 Without RGDS, any viable VICs retained rounded morphology and clumped together rather than spread on the substrate (Supporting Information, Figure S2A,B). Therefore, all hydrogel formulations used for encapsulation contained 2 mM of PEG-RGDS for integrin binding.…”

“…Acryloyl-PEG-RGDS (PEG-RGDS) and acryloyl-PEG-PQ-PEG-acryloyl (PEG-PQ) were conjugated as previously described. 25 Briefly, 3400 Da acryloyl-PEG-succinimidyl valerate (PEG-SVA, Laysan Bio, Arab, AL) was reacted with RGDS at 1:1.2 ratio or PQ at 2.1:1 ratio overnight at pH 8.0. The products were dialyzed for 48 h using a 3500 Da molecular weight cutoff dialysis membrane.…”

Synthetic hydrogels mimicking basement membrane matrices to promote cell-matrix interactions

“…Presentation of RGD on synthetic matrices supports cell adhesion by targeting integrins, such as α v β 3 . Many studies have demonstrated successful BM-like bioactivity of RGD-functionalized hydrogels, as compared to non-functionalized matrices, by directing essential cellular processes such as attachment and spreading (Schmedlen et al, 2002), migration and invasion (Puperi et al, 2015; Halstenberg et al, 2002), and stem cell support (Nuttelman et al, 2005) and differentiation (Saha et al, 2007). Furthermore, other peptides like IKVAV and YIGSR, which are derived from laminin, can also be incorporated into hydrogels together with RGD to orchestrate ligand density-dependent signal presentation independently of the mechanical characteristics of the matrix (Saha et al, 2007; Fittkau et al, 2005).…”