Matricellular Protein Expression and Cell Ultrastructure as Parameters to Test In Vitro Cytotoxicity of a Biomimetic ScaffoldSandra Durante1, Gabriella Teti1, Viviana Salvatore1, Stefano Focaroli1, Monica Mattioli-Belmonte2, Concetta Ferretti2, Adriana Bigi3,
Roberto Giardino4 and Mirella Falconi1*
- *Corresponding Author:
- Mirella Falconi
Department of Biomedical and Neuromotor Sciences (DIBINEM)
University of Bologna, via Irnerio, 48-40126 Bologna (Italy)
Tel: +39 051 2091511
Fax: +39 051 251735
E-mail: [email protected]
Received Date: June 03, 2014; Accepted Date: July 11, 2014; Published Date: July 13, 2014
Citation: Durante S, Teti G, Salvatore V, Focaroli S, Mattioli-Belmonte M, et al. (2014) Matricellular Protein Expression and Cell Ultrastructure as Parameters to Test In Vitro Cytotoxicity of a Biomimetic Scaffold. J Cytol Histol 5:262. doi:10.4172/2157-7099.1000262
Copyright: © 2014 Durante S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Following scaffold implantation, cell sufferance, in-vivo encapsulation, foreign body reaction and inflammatory response has been reported and the up- regulation of matricellular proteins is often connected with this condition. Cytotoxicity of biomaterials is generally tested according to ISO standard 10993-5 based mainly on viability tests. Additional assays, based on improved cytotoxicity knowledge, are suggested in order to better analyze the biocompatibility of implant materials. The purpose of the study was to evaluate the matricellular protein expression as biomarker for in vitro-testing the biocompatibility of implant materials. Tenascin-C, osteocalcin and osteopontin belong to the matricellular protein family and were chosen as cytotoxicity markers. Mesenchymal stem cells were seeded on collagen/hydroxyapatite scaffold and on carboxymethyl cellulose based hydrogel in order to evaluate gene/protein expression by cell viability test, Real Time PCR and western blot. Electron microscopy was carried out to evaluate the morphological changes induced by cell/scaffold interactions. A low expression of tenascin-c, osteonectin and osteopontin was demonstrated in collagen/hydroxyapatite scaffold compared to the cells cultured on tissue flasks and on hydrogel scaffold. Based on our results, we propose matricellular protein expression as parameter for testing in vitro biocompatibility of implant materials.