Fonseca et. al. Enzymatic, physicochemical and biological properties of MMP-sensitive alginate hydro

3d cultures was eectively active in cleaving pvglig

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Unformatted text preview: culture in MMP-sensitive vs. MMP insensitive matrices. Even if differences were not statistically signicant, the same trend has been consistently found between different experiments. However, to better address the effect of MMP-sensitive hydrogels in hMSC differentiation, further studies will have to be carried out to This journal is ª The Royal Society of Chemistry 2013 View Article Online Paper evaluate the expression of more specic osteogenic markers using quantitative assays. Moreover, as observed in 2D, hMSC primarily secreted MMP-2 according to gelatine-zymography data. Importantly, the study showed that although the amounts of secreted MMP-2 decreased along the culture in OM (as in 2D), hMSC in MMP-sensitive PVGLIG/RGD alginate hydrogels expressed higher amounts of MMP-2 than hMSC in control matrices. Downloaded by State University of New York at Buffalo on 16/04/2013 03:56:51. Published on 08 February 2013 on http://pubs.rsc.org | doi:10.1039/C3SM27560D 5 Conclusions This study focused on the characterization of MMP-sensitive alginate hydrogels in terms of their enzymatic, physico-chemical and biological properties. The detailed analysis of the MMP specicity of PVGLIG peptides, and the prospective screening of MMP expression in hMSC cultures, were essential to better elucidate the role of different MMPs in the degradation of the proposed MMP-sensitive hydrogels. In fact, MMP-2 and MMP14 appeared to be key protagonists in that process, at least in vitro. In 3D-cultures, MMP-sensitive alginate hydrogels promoted cell–matrix and cell–cell interactions, and stimulated the secretion of proteases (most likely MMP-2) by hMSC, both in basal and osteogenic conditions. This feature may be advantageous when using MMP-sensitive alginate hydrogels as vehicles for hMSC delivery in tissue regeneration applications. Presumably, these materials will not only provide a more physiological environment to the transplanted cells, but also facilitate their outward migration into the surrounding host tissues, where they will be able to participate more actively in the regeneration process. Acknowledgements This work was nanced by FEDER funds through the Programa Operacional Factores de Competitividade – COMPETE and by ~ Portuguese funds through FCT – Fundaçao para a Ciˆncia e a e Tecnologia, in the framework of the projects Pest-C/SAU/LA0002/ 2011 and BIOMATRIX (PTDC/SAU-BEB/101235/2008 and FCOMP-01-0124-FEDER-010915). KF, FC and CB acknowledge FCT for SFRH/BD/30057/2006, INEB/Unidade 174/BII-01/2008 and Ciˆncia 2008 program, respectively. FRM acknowledges INL e for her PhD scholarship. The authors are grateful to Maria J. Oliveira from INEB for her help with the qRT-PCR analysis and the manuscript revision, to Daniela Silva from CEMUP (Centro de Materiais da Universidade do Porto) for the CryoSEM analysis, and to Paula Gameiro and Tivadar Mach (FCUP) for their support to use of the uorometer. References 1 F. Munarin, P. Petrini, M. C. Tanzi, M. A. Bar...
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This document was uploaded on 09/21/2013.

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