TY - JOUR

T1 - Improving protein delivery of fibroblast growth factor-2 from bacterial inclusion bodies used as cell culture substrates

AU - Seras-Franzoso, Joaquin

AU - Peebo, Karl

AU - García-Fruitós, Elena

AU - Vázquez, Esther

AU - Rinas, Ursula

AU - Villaverde, Antonio

N1 - Funding Information: We are indebted to MINECO ( BFU2010-17450 ), AGAUR ( 2009SGR-0108 ) and CIBER de Bioingeniería, Biomateriales y Nanomedicina (CIBER-BBN, Spain) for funding our research on inclusion bodies. 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. We also acknowledge technical assistance from the Servei de Cultius Cellulars, Producció d’Anticossos i Citometria (SCAC) at the Universitat Autònoma de Barcelona (UAB). A.V. received an ICREA ACADEMIA award. We are also indebted to the Protein Production Platform (CIBER-BBN) for helpful technical assistance and for protein production and purification services ( http://www.bbn.ciber-bbn.es/programas/plataformas/equipamiento ).

PY - 2014/3

Y1 - 2014/3

N2 - Bacterial inclusion bodies (IBs) have recently been used to generate biocompatible cell culture interfaces, with diverse effects on cultured cells such as cell adhesion enhancement, stimulation of cell growth or induction of mesenchymal stem cell differentiation. Additionally, novel applications of IBs as sustained protein delivery systems with potential applications in regenerative medicine have been successfully explored. In this scenario, with IBs gaining significance in the biomedical field, the fine tuning of this functional biomaterial is crucial. In this work, the effect of temperature on fibroblast growth factor-2 (FGF-2) IB production and performance has been evaluated. FGF-2 was overexpressed in Escherichia coli at 25 and 37 C, producing IBs with differences in size, particle structure and biological activity. Cell culture topographies made with FGF-2 IBs biofabricated at 25 C showed higher levels of biological activity as well as a looser supramolecular structure, enabling a higher protein release from the particles. In addition, the controlled use of FGF-2 protein particles enabled the generation of functional topographies with multiple biological activities being effective on diverse cell types.

AB - Bacterial inclusion bodies (IBs) have recently been used to generate biocompatible cell culture interfaces, with diverse effects on cultured cells such as cell adhesion enhancement, stimulation of cell growth or induction of mesenchymal stem cell differentiation. Additionally, novel applications of IBs as sustained protein delivery systems with potential applications in regenerative medicine have been successfully explored. In this scenario, with IBs gaining significance in the biomedical field, the fine tuning of this functional biomaterial is crucial. In this work, the effect of temperature on fibroblast growth factor-2 (FGF-2) IB production and performance has been evaluated. FGF-2 was overexpressed in Escherichia coli at 25 and 37 C, producing IBs with differences in size, particle structure and biological activity. Cell culture topographies made with FGF-2 IBs biofabricated at 25 C showed higher levels of biological activity as well as a looser supramolecular structure, enabling a higher protein release from the particles. In addition, the controlled use of FGF-2 protein particles enabled the generation of functional topographies with multiple biological activities being effective on diverse cell types.

KW - Drug delivery

KW - Inclusion bodies

KW - Nanomedicine

KW - Protein release

KW - Surface functionalization

UR - http://www.scopus.com/inward/record.url?scp=84895068935&partnerID=8YFLogxK

U2 - 10.1016/j.actbio.2013.12.021

DO - 10.1016/j.actbio.2013.12.021

M3 - Article

C2 - 24361427

AN - SCOPUS:84895068935

VL - 10

SP - 1354

EP - 1359

JO - Acta biomaterialia

JF - Acta biomaterialia

SN - 1742-7061

IS - 3

ER -