Abstract
BackgroundMechanobiological studies allow the characterization of cell response to mechanical stresses. Cells need to be supported by a material with properties similar to the physiological environment. Silicone elastomers have been used to produce various in vitro scaffolds of different geometries for endothelial cell studies given its relevant mechanical, optical and surface properties. However, obtaining defined and repeatable properties is a challenge as depending on the different manufacturing and processing steps, mechanical and surface properties may vary significantly between research groups.MethodsThe impact of different manufacturing and processing methods on the mechanical and surface properties was assessed by measuring the Young’s modulus and the contact angle. Silicone samples were produced using different curing temperatures and processed with different sterilization techniques and hydrophilization conditions.ResultsDifferent curing temperatures were used to obtain materials of different stiffness with a chosen silicone elastomer, i.e. Sylgard 184®. Sterilization by boiling had a tendency to stiffen samples cured at lower temperatures whereas UV and ethanol did not alter the material properties. Hydrophilization using sulphuric acid allowed to decrease surface hydrophobicity, however this effect was lost over time as hydrophobic recovery occurred. Extended contact with water maintained decreased hydrophobicity up to 7 days. Mechanobiological studies require complete cell coverage of the scaffolds used prior to mechanical stresses exposure. Different concentrations of fibronectin and collagen were used to coat the scaffolds and cell seeding density was varied to optimize cell coverage.ConclusionThis study highlights the potential bias introduced by manufacturing and processing conditions needed in the preparation of scaffolds used in mechanobiological studies involving endothelial cells. As manufacturing, processing and cell culture conditions are known to influence cell adhesion and function, they should be more thoroughly assessed by research groups that perform such mechanobiological studies using silicone.
Highlights
Mechanobiological studies allow the characterization of cell response to mechanical stresses
The effect on the mechanical and surface properties of the manufacturing and processing techniques commonly used to produce in vitro scaffolds for endothelial mechanobiological studies described in this work can serve as guidelines, highlighting many considerations to take into account
The principal steps involved in manufacturing, as well as the preparation and use of tubular in vitro silicone scaffolds for mechanobiological studies have been examined in order to understand how they truly impact the material properties, cell behavior and the results
Summary
Mechanobiological studies allow the characterization of cell response to mechanical stresses. Silicone elastomers have been used to produce various in vitro scaffolds of different geometries for endothelial cell studies given its relevant mechanical, optical and surface properties. In vivo experiments provide a biological and mechanical environment closer to physiological conditions, it is difficult to study the effect of specific mechanical stresses. In vitro mechanobiological studies are used to expose cells to controlled mechanical stresses in many different conditions in parallel, allowing higher throughput and reduced cost when compared to in vivo studies. In vitro experiments require scaffolds or experimental cell culture models to grow cells in specific geometries, as well as dynamic bioreactors to impose mechanical stresses. These constructs should be adapted to the cell type studied. 3D scaffolds can represent geometrically accurate constructs used to seed an endothelial cell EC monolayer but can represent smooth muscle cells encapsulated into a hydrogel
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