Chethan Kumar1, Basavarajappa Mohana Kumar1, Basan Gowda Kurkalli1, Shishir Shetty2, Shama Rao1, Veena Shetty1. 1. Nitte University Centre for Stem Cell Research and Regenerative Medicine, K. S. Hegde Medical Academy, Nitte (Deemed to Be University), Deralakatte 575018, Mangaluru, India. 2. Department of Conservative Dentistry and Endodontics, A. B. Shetty Memorial Institute of Dental Sciences, Nitte University (Deemed to Be University), Deralakatte 575018, Mangaluru, India.
Abstract
BACKGROUND: A major challenge in bone tissue regeneration is the use of right combination of stem cells with osteoinductive biomaterials. Hence, the present in vitro study was aimed at evaluating the effect of mineralized teeth matrix (MTM) and demineralized teeth matrix (DTM) on the selected cellular and biological characteristics of human dental pulp stem cells (DPSCs). METHODS: Established DPSCs were cultured in conditioned media (CM) of MTM and DTM and analyzed on their morphology, proliferation rate, population doubling time (PDT), viability, migration ability, ploidy and expression of cell surface markers, Further, the effect of MTM and DTM on the biocompatibility and osteogenic differentiation ability of DPSCs was evaluated. RESULTS: The DPSCs exhibited a fibroblast-like morphology with >80% viability. Cells were highly proliferative with an average PDT of 61 ± 12 h. A greater proliferation of DPSCs in the scratched area was observed when cultured in CM of teeth matrix compared to the cells in basal media. Moreover, no chromosomal abnormalities were induced during the culture of DPSCs. Flow cytometry analysis showed that DPSCs in basal media and CM of MTM and DTM were positive for CD29, CD44, CD73, CD90 (>70%), and negative for CD34 and CD45 (<0.1%). Alizarin red staining showed the higher deposition of mineralized nodules in DPSCs cultured with DTM compared to MTM. CONCLUSION: MTM and DTM-derived CM enhanced the proliferation and selected phenotypic markers expression with no chromosomal abnormalities in DPSCs. In addition, both matrices were biocompatible with DPSCs and increased the osteogenic differentiation through higher nodule formation.
BACKGROUND: A major challenge in bone tissue regeneration is the use of right combination of stem cells with osteoinductive biomaterials. Hence, the present in vitro study was aimed at evaluating the effect of mineralized teeth matrix (MTM) and demineralized teeth matrix (DTM) on the selected cellular and biological characteristics of human dental pulp stem cells (DPSCs). METHODS: Established DPSCs were cultured in conditioned media (CM) of MTM and DTM and analyzed on their morphology, proliferation rate, population doubling time (PDT), viability, migration ability, ploidy and expression of cell surface markers, Further, the effect of MTM and DTM on the biocompatibility and osteogenic differentiation ability of DPSCs was evaluated. RESULTS: The DPSCs exhibited a fibroblast-like morphology with >80% viability. Cells were highly proliferative with an average PDT of 61 ± 12 h. A greater proliferation of DPSCs in the scratched area was observed when cultured in CM of teeth matrix compared to the cells in basal media. Moreover, no chromosomal abnormalities were induced during the culture of DPSCs. Flow cytometry analysis showed that DPSCs in basal media and CM of MTM and DTM were positive for CD29, CD44, CD73, CD90 (>70%), and negative for CD34 and CD45 (<0.1%). Alizarin red staining showed the higher deposition of mineralized nodules in DPSCs cultured with DTM compared to MTM. CONCLUSION: MTM and DTM-derived CM enhanced the proliferation and selected phenotypic markers expression with no chromosomal abnormalities in DPSCs. In addition, both matrices were biocompatible with DPSCs and increased the osteogenic differentiation through higher nodule formation.
Authors: Alejandro Victoria-Escandell; José Santiago Ibañez-Cabellos; Sergio Bañuls-Sánchez de Cutanda; Ester Berenguer-Pascual; Jesús Beltrán-García; Eva García-López; Federico V Pallardó; José Luis García-Giménez; Antonio Pallarés-Sabater; Ignacio Zarzosa-López; Manuel Monterde Journal: Stem Cells Int Date: 2017-05-24 Impact factor: 5.443