The nanoscale properties of bacterial inclusion bodies and their effect on mammalian cell proliferation

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).…”

“…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

“…Interestingly, IB physical parameters such as size, wettability, stiffness and superficial charge are within the range of optimal values for surface cell colonization [64]. Moreover, the values of all these features vary between particles produced in bacteria with different genetic background, namely mutants in key genes of the quality control/protein folding machinery, including dnaK, clpA, and clpP [64].…”

“…The high cell membrane avidity of bacterial IBs is probably supported by the amphiphilic nature of the material [64], that might promote an early anchorage to facilitate micropinocytosis. IBs are recognized by the mammalian cell sensing machinery and get closely attached to cell membranes quickly after addition of IBs to the culture media.…”

Improving protein delivery of fibroblast growth factor-2 from bacterial inclusion bodies used as cell culture substrates

“…Moreover, IBs have exhibited a high mechanical plasticity being their physicochemical features regulatable by the proper choice of the producing strain's genetic background [5][6][7]. In addition, the particle size and the conformational quality of the IB forming protein can be tuned by modulating operational variables such as culture temperature or harvesting time as well as engineering the protein construct by the addition of distinct peptide linkers in fusion proteins [8][9][10].…”