Polymodal Activation and Desensitization of TRPV1 Receptor in Human Odontoblasts-Like Cells with Eugenol

Figure 1

Phenotypic characterization of hDPSCs and morphological cell appearance during the cell differentiation process. (a) The PDT was calculated considering the cell count performed in the first 96 hours (4 days) of cell culture, using a hemocytometer. Results are shown as the average of three independent experiments in triplicate (n = 9). (b) The undifferentiated mesenchymal phenotype was verified using flow cytometry by using the cocktail for phenotyping of Miltenyi, which demonstrated positivity for mesenchymal markers CD90, CD105, and CD73 and negativity for the early hematopoietic markers CD14, CD20, CD34, and CD45. A total of 100,000 events were acquired with the use of the FACSCalibur cytometer, and the data were analyzed using FCS Express software. (c) To evaluate the differentiation potential of hDPSCs, they were exposed to differentiation media for the three lineages for 21 days and the respective stains were performed in order to corroborate the acquired phenotype. The osteoblasts were stained with alizarin red, and the formation of mineralization nodules is evidenced. The chondrocytes were stained with Alcian blue, positive labeling of proteoglycans is evidenced, and adipocytes were stained with OilO red; a positive staining of fatty acids is shown. Scale bar: 100 μm. (d) Phase contrast microscopy images showing hDPSCs with tapered, fibroblastic morphology, as well as hOLCs cells during the differentiation process at 7, 14, and 21 days. A trend toward organized training in parallel lines is shown. Leica DM2500 Microscope (Leica Microsystems; Wetzlar, Germany) was used. Scale bar: 100 μm.

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