Biophysical properties of dermal building-blocks affect extra cellular matrix assembly in 3D endogenous macrotissue

Abstract

The fabrication of functional tissue units is one of the major challenges in tissue engineering due to their in vitro use in tissue-on-chip systems, as well as in modular tissue engineering for the construction of macrotissue analogs. In this work, we aim to engineer dermal tissue micromodules obtained by culturing human dermal fibroblasts into porous gelatine microscaffold. We proved that such stromal cells coupled with gelatine microscaffolds are able to synthesize and to assemble an endogenous extracellular matrix (ECM) resulting in tissue micromodules, which evolve their biophysical features over the time. In particular, we found a time-dependent variation of oxygen consumption kinetic parameters, of newly formed ECM stiffness and of micromodules self-aggregation properties. As consequence when used as building blocks to fabricate larger tissues, the initial tissue micromodules state strongly affects the ECM organization and maturation in the final macrotissue. Such results highlight the role of the micromodules properties in controlling the formation of three-dimensional macrotissue in vitro, defining an innovative design criterion for selecting tissue-building blocks for modular tissue engineering.

3D stromal tissue equivalent affects intestinal epithelium morphogenesis in vitro

“…By using tumor-derived cells, such as CaCo-2, a physiological cross-talk was established on 3D-CSSE able to induce CaCo-2 differentiation toward all four highly specialized intestinal cell type that coordinate many physiological functions (absorptive, protective, host defense response). In previous works we have already proved Urciuolo, Imparato, Totaro, & Netti, 2013) that the human dermal fibroblasts are able to synthesize de novo ECM and that the cell-microscaffold interaction triggers mechanotransduction pathways involved in the collagen remodeling (Urciuolo et al, 2016). By partly modified the dynamic culture conditions, in this study we induce the formation of the Intestine-μTP, that are three-dimensional microtissues in which ISEMFs cells are active players in the remodeling of the surrounding scaffold which was gradually substituted by neo-produced ECM components.…”

An Engineered Breast Cancer Model on a Chip to Replicate ECM‐Activation In Vitro during Tumor Progression

“…The optically accessible microfluidic chip hosted two compartments for accommodating both stromal tissue and epithelial tumor tissue, separated by an interface that allowed their physical contact. The 3D stromal tissue consisted of previously engineered tissue micromodules formed by fibroblast‐assembled ECM . We proved that such stroma tissue underwent both cellular and extracellular activation during cancer cell invasion.…”

A three‐dimensional microfluidized liver system to assess hepatic drug metabolism and hepatotoxicity

“…In addition, although spheroid configuration guarantees cell–cell interaction, on the other hand, it does not preserve at all the cell–ECM interplay resulting in a 3D construct in which metabolic activity, mechanical properties, and ECM composition do not resemble the in vivo setting Brancato et al . Among the biomaterials used for this application, gelatin micro‐carries acting as microscaffolds are used in many 3D tissue models (Brancato et al, ; F Urciuolo et al, ; Francesco Urciuolo, Imparato, Totaro, & Netti, ). This type of cell support system provides an ECM‐like structure to the cells and, when used in combination with bioreactor culture, improves cell yield and promotes cell‐matrix interaction (De Gregorio, Imparato, Urciuolo, & Netti, , ; Imparato, Urciuolo, Casale, & Netti, ; Katt, Placone, Wong, Xu, & Searson, ; Nibourg et al, ; Rafiq, Coopman, Nienow, & Hewitt, ; Tan et al, ).…”