In vitro evaluation of dental pulp stem cells (DPSCs) differentiation to odontogenic lineage facilitated by electrospun membranes
By: Dr. Soumya Sheela Aravind
Post-Doctoral Research Associate, Dental Biomaterials Research Group, Sharjah Institute for Medical Research, University of Sharjah
here to join the meeting”
Dental Pulp Stem Cells (DPSCs) are gaining much attention nowadays in the regenerative endodontics field owing to their striking similarity with bone marrow-derived mesenchymal stem cells. The tissue source for in vitro isolation of DPSCs is usually from intact permanent third molars or impacted third molars. The availability of these pulp tissues is certainly a limitation for DPSC culture for pulp-dentin regenerative work. Therefore, researchers have investigated the feasibility of utilizing pulp tissues harvested from patients with irreversible pulpitis to be the tissue source for dental stem cell culture. Owing to the immense therapeutic potential of DPSCs, it is necessary to identify the best isolation method, which provides good quality stem cells that in turn can be utilized for regenerative medicine application. Hence, the present study is an attempt to explore the feasibility of using outgrowth (OG) and enzyme-mediated (COL) digestion protocol in isolating DPSCs from inflamed pulp tissue and its subsequent differentiation potential into the odontogenic lineage. The isolated cells were expanded in culture and the phenotypic characterization was done using flow cytometry. MTT and Real-time PCR assessed the metabolic activity and odontogenic differentiation of the isolated cells. The results showed that the isolated cells were positive for MSC markers such as CD-90, CD-105, and CD-73 and negative for CD-14, CD-45 and STRO-1. MTT assay indicated that the DPSCs from the OG method showed higher metabolic activity than cells from COL. However, the odontogenic differentiation was in favor of cells isolated by the COL method. After confirming that the isolation method and culture conditions do affect the differentiation capacity of isolated cells, the feasibility of utilizing these cells in dental pulp regeneration was also carried out in the latter part of the study. For that, electrospun polycaprolactone (PCL) membranes blended with hydroxyapatite (HA) were developed and the potential of the isolated DPSCs differentiation into odontoblastic lineage was evaluated. Electrospun nanofibrous membrane consisting of PCL blended HA were fabricated and the characterization was done by Scanning electron microscopy (SEM), Fourier- transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis. Cytocompatibility, cell adhesion, and odontogenic differentiation ability of the membranes were assessed by MTT, Live/Dead, SEM/DAPI, qPCR, and SEM-EDS. Among the membranes tested, PCL10%HA showed positive cell attachments, upregulated expression of DSPP, ALP genes, and mineralization. Hence, PCL10%HA proved its potential to be used as a pulp capping material on vital pulp exposure.
Dr. Soumya Sheela Aravind joined as a Postdoctoral Research Associate in 2018 in the Dental Biomaterials Research Group led by Prof. Mehmet Omer Gorduysus at Research Institute of Medical & Health Sciences (RIMHS). She obtained her PhD from Amrita University, India and was working as a Research associate in Le Comité français d'accréditation (COFRAC) tissue culture research lab in India prior to joining UOS. Her research interests include developing different biomaterials and utilizing it for stem cell based regenerative medicine application. She has co-authored seven research papers and two conference proceedings to her credit since her joining at the University of Sharjah.