Two-Dimensional Microscale Engineering of Protein-Based Nanoparticles for Cell Guidance

Abstract

Cell responses, such as positioning, morphological changes, proliferation, and apoptosis, are the result of complex chemical, topographical, and biological stimuli. Here we show the macroscopic responses of cells when nanoscale profiles made with inclusion bodies (IBs) are used for the 2D engineering of biological interfaces at the microscale. A deep statistical data treatment of fibroblasts cultivated on supports patterned with green fluorescent protein and human basic fibroblast growth factor-derived IBs demonstrates that these cells preferentially adhere to the IB areas and align and elongate according to specific patterns. These findings prove the potential of surface patterning with functional IBs as protein-based nanomaterials for tissue engineering.

Production of functional inclusion bodies in endotoxin-free Escherichia coli

“…However, we wanted to ensure that these particles still retained their ability to (i) mechanically stimulate the growth of mammalian cells when used as surface-decorating topographies in cell culture settings as described previously (García-Fruitós et al 2009;Seras-Franzoso et al 2013c;Diez-Gil et al 2010;Tatkiewicz et al 2013;Seras-Franzoso et al 2013b), and (ii) release functional proteins when internalized by mammalian cells (Liovic et al 2012;Vazquez et al 2012;Seras-Franzoso J et al 2013;Seras-Franzoso et al 2013b). The comparative analysis of cell proliferation on IB-decorated surfaces revealed similar properties of all tested IBs (Figure 7).…”

“…Due to their mechanical stability, the ability to penetrate mammalian cells in the absence of cellular damage and the release of functional protein, in a way similar to the release of functional hormones from amyloid repositories (Villaverde 2012), bacterial IBs became unexpectedly promising materials in drug delivery and in regenerative medicine Liovic et al 2012;Talafova et al 2013;García-Fruitós et al 2009;Seras-Franzoso et al 2012;Seras-Franzoso J et al 2013;Seras-Franzoso et al 2013c;Seras-Franzoso et al 2013b;Seras-Franzoso et al 2013a;Tatkiewicz et al 2013;Vazquez et al 2012;Villaverde et al 2012;Villaverde 2012). Although not determined quantitatively, contamination of IBs with bacterial LPS is a result of cell debris formation during cell disruption and separation (Neubauer et al 2006;Georgiou and Valax 1999).…”

“…Bacterial IBs show high biological activity (Garcia-Fruitos et al 2005;GonzalezMontalban et al 2007;Garcia-Fruitos et al 2012), spontaneous penetrability in mammalian cells ) and also mechanical and biological stability García-Fruitós et al 2009), which has allowed to adapt them as nanostructured materials for surface decoration in tissue engineering (García-Fruitós et al 2009;Seras-Franzoso et al 2013b;Tatkiewicz et al 2013;Diez-Gil et al 2010;Seras-Franzoso et al 2011;Seras-Franzoso et al 2012;Seras-Franzoso J et al 2013) and in the context of top-down and bottom-up protein replacement/delivery cell therapies Cano-Garrido et al 2013;Talafova et al 2013;Liovic et al 2012). However, these applications require the bacterial product being free of E.…”

Bacterial Inclusion Bodies: Discovering Their Better Half

“…Also, topographical microscale patterns made of IBs impact on cell positioning, morphology and migration [63]. Interestingly, IB physical parameters such as size, wettability, stiffness and superficial charge are within the range of optimal values for surface cell colonization [64].…”

Integrating Mechanical and Biological Control of Cell Proliferation Through Bioinspired Multieffector Materials

“…IBs show a positive impact on colonization and proliferation (28,29) , and being highly bioadhesive materials, cell expansion on IB-decorated surfaces has been proven to be synergistically supported by both favored adhesion and mechanical stimulation of cell division (30) . In micropatterned surfaces, cells preferentially adhere to IB-rich areas, aligning and elongating according to the IB pattern and choosing the shortest way to reach new adhesion spots on the IBs (31) . Such 2D engineering technique fills the gap between existing techniques which are based on the local modification of the chemical nature of the surface and those based on the modification of the topography at the nanoscale level by physical methods because IBs combine at the same time biofunctionalization and topographical modification of the roughness, as discussed in more detail above.…”