Cytoskeleton and major extracellular matrix proteins were analyzed by immunofluorescence.

<p>Actin was observed in DPSC (A), DPSC with BD (B) and in DPSC with BR (C); vimentin and tubulin (D, E and F); and type 1 collagen and fibronectin (G, H and I). The observed decrease in type 1 collagen, when DPSC were in contact with biomaterials, was confirmed using RT-qPCR (5J), whereas protein secretion was not modified by the experimental conditions (Fig 5K). Scale Bar: 100μm.</p

Media used in the study.

<p>Media used in the study.</p

Composition and properties of the biomaterials.

<p>Composition and properties of the biomaterials.</p

Apoptosis and cell death in contact with the biomaterials.

<p>Detection of apoptosis and cell death. Annexin V and propidium iodide fluorescent stainings of the GSC cultured with no materials (Fig 3A), Enamic (Fig 3B), EHM (Fig 3C), EHMi (Fig 3D) and Z100 (Fig 3E). BCA measurement of the protein concentration in the respective culture conditions (Fig 3F). Flow cytometry analysis of the apoptosis marker Annexin V (Fig 3G) and of dead cells 7-AAD (Fig 3H). Scale bar 100μm.</p

Cytoskeleton and extracellular matrix components analysis.

<p>Actin and Dapi fluorescent stainings of GSC cultured with either Enamic (Fig 4A) or EHM (Fig 4B) or EHMi (Fig 4C) or Z100 (Fig 4D). Tubulin, Vimentin and Dapi immunofluorescent stainings of DPSC cultured with either Enamic (Fig 4E) or EHM (Fig 4F) or EHMi (Fig 4G) or Z100 (Fig 4H). ProCollagen 1alpha1, Fibronectin and Dapi immunofluorescent stainings of GSC cultured with either Enamic (Fig 4I) or EHM (Fig 4J) or EHMi (Fig 4K) or Z100 (Fig 4L). Scale bar 100μm.</p

Primer sequences used for the study.

<p>Primer sequences used for the study.</p

Biomaterials and stem cells characteristics.

<p>SEM microphotographs of the powdered materials. Enamic (Fig 1A), EHM (Fig 1B), EHMi (Fig 1C) and Z100 (Fig 1D). FACS analysis of classical mesenchymal stem cells markers (CD29, 90, 105, 146 and STRO1), of hematopoietic cells (CD45) and of endothalium (CD31) for GSC (Fig 1E) and DPSC (Fig 1F). CFU-F assays of the GSC (Fig 1G). Differentiation staining of the GSC after 14 days: Oil Red O staining (Fig 1H) and Alizarin Red S staining (Fig 1I).Scale bar 100μm.</p

Proliferation and cytoxicity analysis.

<p>Calcein AM viability assays for GSC cultured with the tested materials (Fig 2A-E). Cells microscopic morphology in the same conditions (Fig 2F-J). Scale bar 100μm. Figure 2K: Cellular proliferation analysis. Ki67 indexes graph for the 2 first days of culture in direct contact condition; MTT assays in direct contact or indirect contact for 21 days,. MTT cytotoxicity assays in direct and indirect contact conditions (Fig 2L).</p