Objectives: In this study we evaluated the role of VEGF-A165 and CD90 peptides on proliferation and differentiation of osteoblast-like cells from human Dental Papilla Stem Cells (hDPaSCs). Methods: hDPaSCs were selected after dental bud extraction in 20 young patients (11-15 years) during orthodontic treatment. A stem cells cluster was analyzed with light microscope and SEM before and after confluence and was selected in homogeneous mesenchymal pool with FACS analysis markers (CD29, CD90, CD146, CD166 and STRO-1). Cell samples have been cultured in osteogenic Dulbecco modified medium and the differentiation was evaluated with Alizarin Red and Red Oil. Before and after differentiation, the presence of receivers and co-receivers VEGF-A165 receptors was evaluated in the hDPSCs with RT-PCR, proliferation test, Western Blot for ALP and PECAM-1 and FACS analysis. Results: Cells isolated from dental papilla showed a homogenous cell expressing mesenchymal markers. RT-PCR on undifferentiated hDPSCs highlights the presence of cellular receptors VEGFR-1, VEGFR-2, VEGFR-3, NRP-1 and NRP-2 for the VEGF-A165. The proliferation test showed an increase to 8 days of 70% of the proliferation speed in the differentiated cells dealt with 40ng/ml of VEGF-A165. Western Blot showed an increase of ALP and the absence of PECAM-1. FACS analysis showed a decrease of CD90, not associated with variations of the other investigated mesenchymal markers. Conclusion: The ALP increase and the absence of PECAM-1 confirmed the differentiation in osteogenic sense and not in hematopoietic sense. The CD90 decrease suggests that the VEGF increase the proliferation mechanism and not reduce the differentiation processes. Moreover, VEGF-A165 does not induce shift towards an endothelial phenotype in hDPaSCs, suggesting in stimulating cellular proliferation during hDPaSCs differentiation into osteoblast-like cells, as confirmed from elevated levels of ALP expression. Then, VEGF determine an increase of cell proliferation and differentiation and the results appear in relationship with its concentration
VEGF-A165 AND CD-90 EVALUATION DURING DENTAL PAPILLA STEM CELLS DEVELOPMENT
VINCI , RAFFAELE;GHERLONE , FELICE ENRICO;
2010-01-01
Abstract
Objectives: In this study we evaluated the role of VEGF-A165 and CD90 peptides on proliferation and differentiation of osteoblast-like cells from human Dental Papilla Stem Cells (hDPaSCs). Methods: hDPaSCs were selected after dental bud extraction in 20 young patients (11-15 years) during orthodontic treatment. A stem cells cluster was analyzed with light microscope and SEM before and after confluence and was selected in homogeneous mesenchymal pool with FACS analysis markers (CD29, CD90, CD146, CD166 and STRO-1). Cell samples have been cultured in osteogenic Dulbecco modified medium and the differentiation was evaluated with Alizarin Red and Red Oil. Before and after differentiation, the presence of receivers and co-receivers VEGF-A165 receptors was evaluated in the hDPSCs with RT-PCR, proliferation test, Western Blot for ALP and PECAM-1 and FACS analysis. Results: Cells isolated from dental papilla showed a homogenous cell expressing mesenchymal markers. RT-PCR on undifferentiated hDPSCs highlights the presence of cellular receptors VEGFR-1, VEGFR-2, VEGFR-3, NRP-1 and NRP-2 for the VEGF-A165. The proliferation test showed an increase to 8 days of 70% of the proliferation speed in the differentiated cells dealt with 40ng/ml of VEGF-A165. Western Blot showed an increase of ALP and the absence of PECAM-1. FACS analysis showed a decrease of CD90, not associated with variations of the other investigated mesenchymal markers. Conclusion: The ALP increase and the absence of PECAM-1 confirmed the differentiation in osteogenic sense and not in hematopoietic sense. The CD90 decrease suggests that the VEGF increase the proliferation mechanism and not reduce the differentiation processes. Moreover, VEGF-A165 does not induce shift towards an endothelial phenotype in hDPaSCs, suggesting in stimulating cellular proliferation during hDPaSCs differentiation into osteoblast-like cells, as confirmed from elevated levels of ALP expression. Then, VEGF determine an increase of cell proliferation and differentiation and the results appear in relationship with its concentrationI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
