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Abstract
Background: Bacterial inclusion bodies (IBs), mechanically stable, submicron protein particles of 50–500 nm dramatically favor mammalian cell spread when used for substrate surface decoration. The mechanisms supporting fast colonization of IB-modified surfaces have not yet been identified. Results: This study provides evidence of mechanotransduction-mediated stimulation of mammalian cell proliferation on IB-decorated surfaces, as observed by the enhanced phosphorylation of the signal-regulated protein kinase and by the dramatic emission of filopodia in the presence of IBs. Interestingly, the results also show that IBs are highly bioadhesive materials, and that mammalian cell expansion on IBs is synergistically supported by both enhanced adhesion and mechanical stimulation of cell division. Discussion: The extent in which these events influence cell growth depends on the particular cell line response but it is also determined by the genetic background of the IB-producing bacteria, thus opening exciting possibilities for the fine tailoring of protein nanoparticle features that are relevant in tissue engineering.
Original submitted: 7 February 2011; Revised submitted: 7 April 2011
Financial & competing interests disclosure
A Villaverde, E García–Fruitós, E Vazquez, J Veciana, I Ratera and C Díez-Gil are coinventors of a patent (P200900045) on the use of inclusion bodies as reagents for mammalian cell culture. This study has been funded by MICINN (BFU2010-17450, CTQ2006-06333 and CTQ2010-19501), AGAUR (2009SGR-00108 and 2009SGR-00516) and CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN, Spain). CIBER-BBN is an initiative funded by the VI National R&D&i Plan 2008–2011, Iniciativa Ingenio 2010, Consolider Program, CIBER Actions and financed by the Instituto de Salud Carlos III with assistance from the European Regional Development Fund. Joaquin Seras-Franzoso is recipient of a PIF doctoral fellowship from UAB, and Antonio Villaverde of an ICREA ACADEMIA award. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.
No writing assistance was utilized in the production of this manuscript.
Ethical conduct of research
The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.
Acknowledgements
The authors are indebted to the Cell Culture Unit of the Servei de Cultius Cellulars, Producció d‘Anticossos i Citometria (SCAC), and to the Servei de Microscòpia, both at the UAB. The authors thank the Protein Production Platform (CIBER-BBN) for helpful technical assistance and for protein production and purification services (http://bbn.ciber-bbn.es/programas/plataformas/equipamiento). The authors also appreciate helpful discussions with M Dalby about ERK biology and valuable advice from JM Lizcano and his group.