In Vitro Cultivation, Characterization and Osteogenic Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth on 3D Printed Polylactic Acid Scaffolds

Tissue engineering mainly focuses on creating appropriate conditions for the regeneration of tissues. Scaffolds, signal molecules, and stem cells interact with each other and compose the essential components of this field.
This study aimed at investigating the osteogenic induction ability of PLA Poly Lactic Acid (PLA) scaffolds and comparing the osteogenic differentiation behavior of Stem Cells from Human Exfoliated Deciduous Teeth (hSHEDs) in standard culture medium and on PLA scaffolds.
The current clinical experimental study was conducted between April 2016 and October 2016 at the Near East University cell culture laboratory located in North Cyprus. The pulp tissues of deciduous teeth (non-decayed and in the absence of abscess, fistula or periapical lesion) were sampled from 10 healthy children aged between 6 and 11 years. The isolated hSHEDs were divided to 4 groups. The control group/Group1 consisted of cells, which were cultivated in standard culture medium, and Group2 cells were differentiated into an osteogenic lineage using osteogenic differentiation medium. Group 3 represented the non-differentiated group, which was transferred onto three dimensional (3D) printed PLA scaffolds and Group 4 cells were differentiated to the osteogenic lineage and transferred onto 3D printed PLA scaffolds. All groups were analyzed immunohistochemically and by immune-labeling, and were evaluated semi-quantitatively using the HSCORE.
Cultivation of hSHEDS on PLA scaffolds was assessed for 14 and 21 days; osteogenic differentiation was detected both histochemically and immunohistochemically. Generally, Osteocalcin (OCN) immunoreactivities were higher than Osteonectin (ON) immunoreactions in all groups. Despite higher OCN immunoreactivities, the intensities of OCN between 14 days and 21 days in group 4 (497.3 ± 0.57% and 486.7 ± 5.77%, respectively) were similar (P > 0.05). While the intensity of ON was 280.0 ± 10% in group 4, in group 2 the intensity of ON was 206.7 ± 5.77%, and on the 14th day the results were statistically significant (P Poly lactic acid is a suitable scaffold material for osteogenic induction of the hSHEDs. The expression patterns of both markers showed that a 14-day cultivation period is adequate for hSHEDs with/without PLA scaffolds to differentiate into osteoblasts.