Objective: The need to restore and maintain the function of human bone tissue in skeletal defects makes bone tissue engineering a valuable strategy for tissue regeneration, thanks to the combination between appropriate materials and mesenchymal stem cells. Aim of this study was to evaluate the proliferation and mineralization of human adipose-derived stem cells (hADSCs) in association with poli(Ɛ-caprolactone) 3D scaffold (3D-PCL) for future application in regenerative medicine. Material and Methods: hADSCs were cultured in growth medium (10% FBS, 1 ng/ml bFGF and 1% antibiotics) at 37°C and 5% CO2. hADSCs were plated on 3D-PCL to evaluate fibronectin adhesion protein and cytoskeleton by immunocytochemistry. Proliferation was analyzed by MTT assay after cell seeding on 3D-PCL at appropriate density. Alkaline Phospatase (ALP) activity and mineralization were quantified, during osteoinduction, by spectrofluorometric assays. Statistical analysis was performed by linearity test and Student’s t-test, comparing the values obtained on 3D-PCL with those obtained on PCL film, used as control [1]. Results: No qualitative differences were found in cell adhesion and cytoskeleton fiber morphology between hADSCs grown on 3D-PCL and hADSCs grown on PCL film. MTT assay has measured a consistent growth of absorbance during all the studied period (from 2 to 16 days) for hADSCs seeded on 3D-PCL (linear regression r2= 0,94), suggesting that good cell proliferation and cytocompatibility were achieved on the scaffold. Osteogenic differentiation of hADSCs on 3D-PCL was assessed from 0 to 35 days and has shown not only a significant increase on ALP activity, after 21 days of induction (+ 4579 % vs time 0), but also on Ca2+ deposition, after 35 days of induction (+ 6859 % vs time 0). The same and not significantly different behavior was obtained with hADSCs induced on film PCL, as control. Conclusion: This work has shown encouraging results in terms of proliferation and mineralization of hAMSCs on 3D-PCL scaffold prepared by wet spinning, suggesting and promoting the research of new strategies to improve this model for future applications in bone tissue regeneration. Reference: [1] Romagnoli C. et al. Biomed Res Int. 2015. Acknowledgments: Regione Toscana-POR CREO FESR 2007-2013. Progetto REOSS.

PROLIFERATION AND MINERALIZATION OF HUMAN MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE ON POLY(Ɛ-CAPROLACTONE) 3D SCAFFOLD FOR BONE TISSUE ENGINEERING APPLICATION / Romagnoli, Cecilia; Zonefrati, Roberto; Galli, Gianna; Puppi, Dario; Pirosa, Alessandro; Chiellini, Federica; Martelli, Francesco Saverio; Tanini, Annalisa; Brandi, Maria Luisa. - In: OSTEOPOROSIS INTERNATIONAL. - ISSN 0937-941X. - ELETTRONICO. - (2016), pp. 0-0.

PROLIFERATION AND MINERALIZATION OF HUMAN MESENCHYMAL STEM CELLS DERIVED FROM ADIPOSE TISSUE ON POLY(Ɛ-CAPROLACTONE) 3D SCAFFOLD FOR BONE TISSUE ENGINEERING APPLICATION

ROMAGNOLI, CECILIA;ZONEFRATI, ROBERTO;GALLI, GIANNA;TANINI, ANNALISA;BRANDI, MARIA LUISA
2016

Abstract

Objective: The need to restore and maintain the function of human bone tissue in skeletal defects makes bone tissue engineering a valuable strategy for tissue regeneration, thanks to the combination between appropriate materials and mesenchymal stem cells. Aim of this study was to evaluate the proliferation and mineralization of human adipose-derived stem cells (hADSCs) in association with poli(Ɛ-caprolactone) 3D scaffold (3D-PCL) for future application in regenerative medicine. Material and Methods: hADSCs were cultured in growth medium (10% FBS, 1 ng/ml bFGF and 1% antibiotics) at 37°C and 5% CO2. hADSCs were plated on 3D-PCL to evaluate fibronectin adhesion protein and cytoskeleton by immunocytochemistry. Proliferation was analyzed by MTT assay after cell seeding on 3D-PCL at appropriate density. Alkaline Phospatase (ALP) activity and mineralization were quantified, during osteoinduction, by spectrofluorometric assays. Statistical analysis was performed by linearity test and Student’s t-test, comparing the values obtained on 3D-PCL with those obtained on PCL film, used as control [1]. Results: No qualitative differences were found in cell adhesion and cytoskeleton fiber morphology between hADSCs grown on 3D-PCL and hADSCs grown on PCL film. MTT assay has measured a consistent growth of absorbance during all the studied period (from 2 to 16 days) for hADSCs seeded on 3D-PCL (linear regression r2= 0,94), suggesting that good cell proliferation and cytocompatibility were achieved on the scaffold. Osteogenic differentiation of hADSCs on 3D-PCL was assessed from 0 to 35 days and has shown not only a significant increase on ALP activity, after 21 days of induction (+ 4579 % vs time 0), but also on Ca2+ deposition, after 35 days of induction (+ 6859 % vs time 0). The same and not significantly different behavior was obtained with hADSCs induced on film PCL, as control. Conclusion: This work has shown encouraging results in terms of proliferation and mineralization of hAMSCs on 3D-PCL scaffold prepared by wet spinning, suggesting and promoting the research of new strategies to improve this model for future applications in bone tissue regeneration. Reference: [1] Romagnoli C. et al. Biomed Res Int. 2015. Acknowledgments: Regione Toscana-POR CREO FESR 2007-2013. Progetto REOSS.
2016
Romagnoli, Cecilia; Zonefrati, Roberto; Galli, Gianna; Puppi, Dario; Pirosa, Alessandro; Chiellini, Federica; Martelli, Francesco Saverio; Tanini, Annalisa; Brandi, Maria Luisa
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/1044609
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