Tuning the Nanotopography and Chemical Functionality of 3D Printed Scaffolds through Cellulose Nanocrystal Coatings

Abstract

In nature, cells exist in three-dimensional (3D) microenvironments with topography, stiffness, surface chemistry, and biological factors that strongly dictate their phenotype and behavior. The cellular microenvironment is an organized structure or scaffold that, together with the cells that live within it, make up living tissue. To mimic these systems and understand how the different properties of a scaffold, such as adhesion, proliferation, or function, influence cell behavior, we need to be able to fabricate cellular microenvironments with tunable properties. In this work, the nanotopography and functionality of scaffolds for cell culture were modified by coating 3D printed materials (DS3000 and poly­(ethylene glycol)­diacrylate, PEG-DA) with cellulose nanocrystals (CNCs). This general approach was demonstrated on a variety of structures designed to incorporate macro- and microscale features fabricated using photopolymerization and 3D printing techniques. Atomic force microscopy was used to characterize the CNC coatings and showed the ability to tune their density and in turn the surface nanoroughness from isolated nanoparticles to dense surface coverage. The ability to tune the density of CNCs on 3D printed structures could be leveraged to control the attachment and morphology of prostate cancer cells. It was also possible to introduce functionalization onto the surface of these scaffolds, either by directly coating them with CNCs grafted with the functionality of interest or with a more general approach of functionalizing the CNCs after coating using biotin–streptavidin coupling. The ability to carefully tune the nanostructure and functionalization of different 3D-printable materials is a step forward to creating in vitro scaffolds that mimic the nanoscale features of natural microenvironments, which are key to understanding their impact on cells and developing artificial tissues.

Two-Photon Polymerization of 2.5D and 3D Microstructures Fostering a Ramified Resting Phenotype in Primary Microglia

“…The most significant limitation of 2PP is the relatively long printing time hindering the upscaling of the technology ( Moroni et al, 2018 ). Multiple polymeric, hydrogel or composite materials were employed in combination with 2PP to fabricate micro- and nano-patterns as well as 3D structures for in vitro cellular studies involving neuroblastoma, glioblastoma, prostate cancer, murine cerebellar granule, chondrocytes, macrophages, neuronal and stem cells ( Marino et al, 2013 ; Accardo et al, 2017 , 2018 ; Turunen et al, 2017 ; Maciulaitis et al, 2019 ; Fendler et al, 2019 ; Babi et al, 2021 ; Bertels et al, 2021 ; Maciulaitis et al, 2021 ; Nouri-Goushki et al, 2021 ; Akolawala et al, 2022 ; Costa et al, 2022 ).…”

3D‐Engineered Scaffolds to Study Microtubes and Localization of Epidermal Growth Factor Receptor in Patient‐Derived Glioma Cells

“…pore size, surface nanotopography, rod size, curvature) on the behavior and morphology of glioma cell can be investigated. [ 38 ]…”

Influence of Trabecular Geometry on Scaffold Mechanical Behavior and MG-63 Cell Viability

“…Several authors in the literature improve the DS3000’s compatibility, reducing the harmful impact of byproducts, by coating the substrate with fibronectin, favoring the hydrophilic behavior of the material surface [ 19 , 25 ]. Other authors, such as Babi et al [ 27 ] used dip coating with cellulose nanocrystals (CNCs) for tuning the nano-topography and functionality of 3D-printed scaffolds in DS3000.…”