ECTS Abstracts (2015) 1 P182

Viability and Profileration of Human Adipose-Derived Stem Cells on Poly(?-Caprolactone) Films for Bone Tissue Engineering Applications

Cecilia Romagnoli1, Roberto Zonefrati1, Elisa Fierini1, Dario Puppi2, Federica Chiellini2, Emo Chiellini2, Annalisa Tanini1, Francesco Saverio Martelli3 & Maria Luisa Brandi1

1University of Florence, Florence, Italy, Italy, 2University of Pisa, Pisa, Italy, Italy, 3Biomolecular Diagnostic Srl, Florence, Italy, Italy.

Bone tissue engineering represents a challenge in regenerative medicine. Many type of materials have been studied as scaffolds in combination with mesenchymal stem cells, in order to develop viable substitutes able to restore and maintain the function of human bone tissues in skeletal defects. In this work, we have evaluated the viability, the adhesion and the proliferation of human adipose-derived stem cells (hADSCs) in association with poli(?-caprolactone), PCL, a biodegradable polyester, for future application in regenerative medicine. After the Local Ethical Committee approval, human adipose tissue was minced and digested at 37 °C in 3 mg/ml collagenase type I, for three hours. Primary cells were cultured in growth medium (10% FBS, 1 ng/ml bFGF and 1% antibiotics) at 37 °C and 5% CO2. Characterized hADSCs, (CD44+, CD105+, STRO1+ and CD45-), were plated on PCL film to evaluate fibronectin adhesion protein and cytoskeleton, by immunocytochemistry, using laser scanning confocal microscopy. Cell viability and proliferation were analysed by Acridine Orange (AO) staining and MTT assay on days 2, 4, 7, 10, 13, 16 after cell seeding on PCL film at appropriate density (5 x 103). Statistical analysis was performed by linearity test. No qualitative differences were found in cell adhesion and cytoskeleton fiber morphology between hADSCs grown on PCL film and hADSCs grown on polystyrene, used as control. MTT assay has measured a consistent growth of absorbance during all the studied period for hADSCs seeded on PCL (linear regression r2=0,86), suggesting that good cell viability was achieved on biomaterial. Cytocompatibility was also confirmed by AO staining microscopic observations. PCL construct has shown encouraging results in terms of cell adhesion and viability of hADSCs, opening new possibilities for future application in bone tissue engineering. Experiments are in progress in order to evaluate the osteogenic differentiation process of hADSCs on PCL.

Disclosure: The authors declared no competing interests. This work was supported by Regione Toscana-POR CREO FESR 2007-2013. BANDO UNICO R&S 2012, linea A. Decreto dirigenziale n.2746, 17/06/2014. Progetto REOSS.