Engineered Fibrillar Fibronectin Networks as Three‐Dimensional Tissue Scaffolds

Abstract

Extracellular matrix (ECM) proteins, and most prominently, fibronectin (Fn), are routinely used in the form of adsorbed pre‐coatings in an attempt to create a cell‐supporting environment in both two‐ and three‐dimensional cell culture systems. However, these protein coatings are typically deposited in a form which is structurally and functionally distinct from the ECM‐constituting fibrillar protein networks naturally deposited by cells. Here, the cell‐free and scalable synthesis of freely suspended and mechanically robust three‐dimensional (3D) networks of fibrillar fibronectin (fFn) supported by tessellated polymer scaffolds is reported. Hydrodynamically induced Fn fibrillogenesis at the three‐phase contact line between air, an Fn solution, and a tessellated scaffold microstructure yields extended protein networks. Importantly, engineered fFn networks promote cell invasion and proliferation, enable in vitro expansion of primary cancer cells, and induce an epithelial‐to‐mesenchymal transition in cancer cells. Engineered fFn networks support the formation of multicellular cancer structures cells from plural effusions of cancer patients. With further work, engineered fFn networks can have a transformative impact on fundamental cell studies, precision medicine, pharmaceutical testing, and pre‐clinical diagnostics.

Enhanced mitochondrial fission suppresses signaling and metastasis in triple-negative breast cancer

“…To study the effects of PISD on cell migration in a complex 3D environment, we used tissue-engineering constructs consisting of a SU8 construct coated with networks of engineered fibrillar fibronectin [24]. We seeded 5 × 10 4 SUM159-CBG-WT or PISD cells stably expressing pHAEP onto the construct while rotating for 1 h at 37°C.…”

The Dynamic Relationship of Breast Cancer Cells and Fibroblasts in Fibronectin Accumulation at Primary and Metastatic Tumor Sites

“…Cells in chamber slides or 6 well plates were decellularized before immunoblotting or immunostaining for FN (Sigma, St. Louis, MI, USA, Cat#3648, 1:200 dilution) when indicated as previously described [6,58]. Starting with live cells, cells were washed with PBS then deionized water.…”

3D printing of tissue engineering scaffolds: a focus on vascular regeneration

“…In contrast, in strategies of printing of biomaterial inks and cell adhesion, the available selection of materials and processes is wider. Biomaterials, such as fibronectin, were reported as a surface-adsorbed pre-coating for scaffolds [ 71 ]. But cells are not easy to locate in 3D space.…”

“…During fabrications of vascular scaffolds, auxiliary processes are used as a complement to the forming processes. There are three aspects usually involved using auxiliary materials: (i) chemical modification [ 78 ] and physical modification [ 75 ] of printing materials; (ii) printing processes [ 22 , 23 , 79 ]; and (iii) post-treatment processes [ 34 , 71 ]. Some materials such as methacrylate, GMA, and magnesium chloride hexahydrate are utilized for chemical modification [ 35 , 78 ].…”