Evaluating Stemness Markers in hDPSCs on Titanium

In order to evaluate the biological properties of hDPSCs seeded on titanium and titanium–zirconia (CERID) disks before and after treatment, stemness markers were analyzed. Particularly, after 72 h of culture, cells were fixed and permeabilized as described above. After blocking with 3% bovine serum albumin (BSA) in PBS, cells were immunolabeled with mouse anti-nestin (Merck Millipore, Burlington, MA, United States) and rabbit anti-SOX10 (Abcam, Cambridge, UK) primary antibodies and subsequently targeted by AlexaFluor-conjugated secondary antibodies (Thermo Fisher Scientific, Waltham, MS, USA) [26 (link)]. A Nikon A1 confocal laser scanning microscope was used to perform confocal imaging. Confocal serial sections were processed with Fiji ImageJ software (Fiji for Mac OS X; NIH, Bethesda, MD, USA) to obtain 3-dimensional projections, and image rendering was conducted with Adobe Photoshop software (Adobe Photoshop 2022, ver. 23.5.2) [26 (link)].