Incorporation of AuNP-PLL nanocomplexes in DPSC: a new tool for 3D analysis in pulp regeneration

To assess the viability of dental pulp stem cells loaded with gold nanoparticles complexed with poly (L-lysine) (AuNP-PLL) and to track the cellular behavior in a 3D analysis by micro-CT.

DPSC (dental pulp stem cells) were cultured and incorporated with AuNP-PLL (0.2 mg/ml) and assessed for cell viability (24 h, 48 h, and 72 h) using MTS assay. Apoptosis/cell death index and cell cycle were analyzed by propidium iodide. AuNP-PLL-RITC were used for observation in confocal microscopy and quantification of the incorporation rates. Cells were also suspended in agarose and analyzed three-dimensionally in μCT, assessing their radiopacity. Quantitative data (cell viability and apoptosis) were analyzed by t test (p < 0.05).

AuNP-PLL labeling did not affect cellular viability in any of the periods analyzed nor interfered with the apoptosis index of DPSC. AuNP-PLL nanocomplexes were identified in the cytoplasm of cells by immunofluorescence, mainly in the perinuclear region. The observed incorporation rate was 98%. Micro-CT analysis has shown that incorporated cells are now visible using x-ray, with a clear increase in radiopacity when compared to the control group (non-incorporated cells).

These results indicate that it is possible to incorporate AuNP-PLL complex into DPSC and track the cells by using μCT; furthermore, this incorporation of 0.2 mg/ml of AuNP-PLL does not interfere in the DPSC basic behavior.

This methodology can be a useful tool for cellular labeling to observe cell behavior and their interaction with scaffolds in a 3D manner, opening an array of new approaches in regenerative endodontics.