Abstract
The aim of the present research was the evaluation of the behavior of human periodontal ligament stem cells (hPDLSCs), cultured in presence of Endobon® Xenograft Granules (G), a fully deproteinated hydroxyapatite ceramic scaffold derived from cancellous bovine bone. hPDLSCs were seeded with and without G for 24 h to 1 week. The cell growth, morphological features, adhesiveness, differentiation ability, modulation of miR-210 and Vascular Endothelial Growth Factor (VEGF) secretion were analyzed by means of MTT assay, Scanning Electron Microscopy (SEM), Confocal Laser Scanning Microscopy (CLSM), Alizarin Red S assay, RT-PCR and ELISA test, respectively. hPDLSCs grown on the biomaterial showed the ability to form focal adhesion on the substrate, as demonstrated by vinculin expression. These data were supported by SEM analysis showing that an adhesiveness process associated to cell growth occurs between cells and biomaterials. The osteogenic differentiation, evaluated by morphological, biochemical, and RT-PCR analysis, was pronounced in the hPDLSCs grown in the three-dimensional inorganic bovine bone substitute in the presence of osteoinductive conditions. In addition, an upregulation of miR-210 and VEGF was evident in cells cultured in presence of the biomaterial. Our results inspire us to consider granules not only an adequate biocompatible three-dimensional biomaterial, but also an effective inductor of miR-210 and VEGF; in fact, the involvement of miR-210 in VEGF secretion could offer a novel regulatory system in the early steps of the bone-regeneration process.
Highlights
Periodontal disease is a wide range of inflammatory diseases leading to bone and tooth loss [1,2]
We have recently reported that stem cells derived from human periodontal ligament can be an easy and efficient autologous source of stem cells with high expansion capacity and the ability to differentiate in osteogenic cells that can colonize and grow connected to a biocompatible scaffold [9,10,11,12,13]
Our results indicate that the 3D granules in contact with human periodontal ligament stem cells (hPDLSCs) showed osteoconductive properties evaluated through adhesion and proliferation, and the ability to stimulate Vascular Endothelial Growth Factor (VEGF) secretion from cells related to miR-210 expression
Summary
Periodontal disease is a wide range of inflammatory diseases leading to bone and tooth loss [1,2]. The essential components needed in bone-tissue engineering for successful results are represented by an appropriate scaffold and a suitable stem-cell source [5,6]. Other important elements that have a key role in the bone-regeneration process are represented by growth and differentiation factors, which, by autocrine/paracrine mechanisms, can mediate reparative and regenerative actions. Among these proteins, Vascular Endothelial Growth Factor (VEGF) is known to act as promoter of angiogenesis [14], which plays a pivotal role in regenerative processes
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