Incorporation of AuNP-PLL nanocomplexes in DPSC: a new tool for 3D analysis in pulp regeneration
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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.
Materials and methods
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.
KeywordsTissue engineering Stem cells Dental pulp Scaffold Endodontics
The authors would like to acknowledge use of the University of Iowa Central Microscopy Research Facility, a core resource supported by the Vice President for Research & Economic Development, the Holden Comprehensive Cancer Center and the Carver College of Medicine.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
For this type of study, ethical approval is not required.
For this type of study, formal consent is not required.
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