Iran Red Crescent Med J. 2017 August; 19(8):e55593. Published online 2017 July 22. doi: 10.5812/ircmj.55593. Brief Report In Vitro Cultivation, Characterization and Osteogenic Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth on 3D Printed Polylactic Acid Scaffolds Aylin Islam, 1,* Emil Mammadov, 2 Remziye Kendirci, 3 Ersin Aytac, 4 Serap Cetiner, 1 and Hafize Seda Vatansever 3,5 1 Paediatric Dentistry Department, Near East University, Nicosia, North Cyprus 2 Paediatric Surgery Department, NEU3D Laboratories, Near East University, Nicosia, North Cyprus 3 Histology and Embriyology Department, Celal Bayar University, Manisa, Turkey 4 NEU3D Laboratories, Near East University, Nicosia, North Cyprus 5 Experimental Health Science Research Center, Near East University, Nicosia, North Cyprus * Corresponding author: Aylin Islam, Near East Boulevard, Near East University, Nicosia/North Cyprus, Mersin 10, Turkey. Tel: +90-3926802030/2607/2635, Fax: +90-3926802025, E-mail: aylin.islam89@gmail.com Received 2017 February 08; Revised 2017 March 02; Accepted 2017 March 21. Abstract Background: Tissue engineering mainly focuses on creating appropriate conditions for the regeneration of tissues. Scaffolds, sig- nal molecules, and stem cells interact with each other and compose the essential components of this field. Objectives: 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. Methods: 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, fis- tula 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 dif- ferentiated 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 lin- eage and transferred onto 3D printed PLA scaffolds. All groups were analyzed immunohistochemically and by immune-labeling, and were evaluated semi-quantitatively using the HSCORE. Results: Cultivation of hSHEDS on PLA scaffolds was assessed for 14 and 21 days; osteogenic differentiation was detected both his- tochemically 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 < 0.0001). Conclusions: 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. Keywords: Cell Differentiation, Dentistry, Stem Cell, Tissue Engineering 1. Background Dental tissue loss due to trauma, disease or congenital abnormalities generates a major health issue worldwide (1). Regenerative medicine is a rapidly developing field and newly emerging fields such as tissue engineering, ma- terial science, cell and molecular biology have been devel- oped, some of which incorporate the use of biotechnolo- gies to provide tissue regeneration (2). 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 essen- tial components of this field (3). Stem cells were quickly adopted as key elements of tissue engineering by means of colonization of self- renewable progenitor cells to constitute one or more cell types. Currently, stem cell contents from dental structures have been examined by various research groups. These ar- eas are pulpal structures of primary and permanent teeth, periodontal ligaments, apical papilla, and dental follicles (4, 5). Three-dimensional (3D) printing is used as a method Copyright © 2017, Iranian Red Crescent Medical Journal. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License (http://creativecommons.org/licenses/by-nc/4.0/) which permits copy and redistribute the material just in noncommercial usages, provided the original work is properly cited.