Study of rMSC viability, morphology and adhesion on different bone mimetic surfaces

NOELIA D' ELÍA; COLLEEN MATHIEU; CAROLINE D. HOEMANN; A. NOEL GRAVINA; JAVIER SARTUQUI; PAULA V. MESSINA

BONE

ELSEVIER SCIENCE INC

Lugar: Amsterdam; Año: 2016 vol. 89 p. 68 - 68

8756-3282

Rabbit mesenchymal stem cells (rMSC) were seeded on different ratios of Hydroxyapatite/Collagen type I (HA/CoI) coating amounts in an attempt to mimic a remodeling bone environment in which they attach and differentiate. HA nanomaterial was composed of nanoparticles with similar physicochemical characteristics of mineral bone matrix. Recently it was shown that the elevation of α-smooth muscle actin (α-SMA) expression is associated with MSC´s osteogenic potential. Also, actin cytoskeleton organization is sensitive to material´s topography. Therefore, during this study MSC´s adhesion and spreading on HA/CoI coatings, with the same inorganic-organic matrix proportion of the bone tissue, was evaluated by α-SMA immunostaining, number of cells attached and MTT assay. Images obtained by 2-photon laser confocal microscopy showed that attached cells had a typical MSC morphology, with a spread polygonal shape and cytoplasmic extensions; also homogeneous cell distribution was found on biomimetic coatings using HA amounts from 15 to 62 μg/cm2 and 31 μg/cm2 of CoI. Semi-quantitative images analysis showed that α-SMA content was the same as the control and MTT assay demonstrated that mitochondrial metabolic activity was not affected (p > 0.10). On the other hand, cells seeded on a hundred times larger HA coating amount showed less spreading and rounded cells, also adhesion and metabolic activity were inhibited (p ˂ 0.01). The obtained results define a maximal MSC cytocompatible HA coating amount. On the whole, results obtained showed that no differences were found on cells viability, cytoskeleton integrity and MSC phenotype on the different biomimetic coating tested.