Yessenia Valverde
University of Illinois at Chicago, USA
Title: PARP-1 inhibition enhances BDNF secretion in dental pulp stem cells derived odontoblast-like cells
Submitted Date: 26/10/2017
Biography
Yessenia Valverde has completed her PhD in 2013 from Niigata University Graduate school of Medical and Dental Sciences, Japan and currently persuing postdoctoralrnresearch from University of Illinois at Chicago College of Dentistry. She has exemplary training in rodent model of dental nerve denervation with the BDNF modulation.
Abstract
The nuclear enzyme poly(ADP-ribose) polymerase (PARP-1) has been implicated its role in several stem cells fate determinationrnand differentiation. The role of PARP-1 in dental pulp stem cell (DPSC) differentiation especially in the context of its ability tornmodulate nerve regeneration factor has not been ivestigated. Neurotrophins are an essential group of nerve regeneration signals. Inrnthis study, we investigated the role of PARP-1 in the modulation of brain-derived neurotrophic factor (BDNF) in DPSCs derivedrnodontoblast-like cells. Human DPSCs were prepared from healthy molars at the 2/3 root formation stage by the explant outgrowthrnmethod. DPSCs were cultured in regular media and osteogenic media and treated with PARP-1 antagonist and PARP1 exogeneousrnprotein for 72 hours in regular media (regular growth media), and then swapped with osteogenic media for 21 days. The PARP-rn1 inhibitor and protein were treated every three days during the whole differentiation process. Immunohistochmistry, PCR andrnwestern analysis for the BDNF and various differentiation markers were performed. Our PCR results demonstrate that differentiatedrncells show odontoblast-like properties as they express odontogenic markers such as DSPP and RUNX. There is possibility thatrnthe PARP1 treatment induces DPSCs into other cell types. Some show very unique morphology with large cytoplasm and ovalrnnucleus. PARP-1 inhibition significantly increased BDNF secretion in the diffentiated cells. This observation was confirmed by bothrnimmunohistochemistry and western blot. Taken together, our results indicate that PARP-1 constitutes a negative regulator of thernBDNF secretion during odontogenic DPSC differentiation demonstrating its potential for successful nerve regeneration engineeringrnstrategies.
Amer Marachli
University of Illinois at Chicago, USA
Title: Complement C5a promotes odontogenic dental pulp stem cell differentiation
Submitted Date: 26/10/2017
Biography
Amer Marachli is an undergraduate student at the University of Illinois at Urbana-Champaign with a bachelors in science in Molecuar and Cellular Biology. He joined Dr.rnSeung Chung`s lab as a Summer Research Intern in June 2017, and successfully completed his project regarding the role of C5a in DPSC odontogenic differentiation. Hernplans to apply to dental schools in the fall of 2017.
Abstract
Dental pulp stem cells (DPSCs) are multipotent/undifferentiated cells that are found in the soft living tissue within the dentalrnpulp. Given their easy access compared to other stem cells, they have been given much attention in recent regenerative medicine.rnOdontoblasts are dentin-making cells in the pulp and have a significant role in dentin-pulp repair in injured teeth. Our teamrndemonstrated for the first time that the complement system, a major component of innate immunity and inflammation, is activatedrnat the injured site of human carious teeth and plays an important role in dental nerve regeneration. We extend this observationrnin DPSC differentiation. The role of complement fragment C5a in the differentiation of DPSCs is unknown. Human DPSCs werernprepared from healthy molars at the 2/3 root formation stage by the explant outgrowth method. DPSCs were cultured in regularrnmedia and osteogenic media and treated with C5a antagonist and C5a exogeneous protein for 72 hours in regular growth media,rnand then swapped with osteogenic media for 21 days.The C5a inhibitor and protein were treated every three days during the wholerndifferentiation process. Immunohistochmistry, PCR and western analysis for various differentiation markers were performed. C5arnplays a positive role in odontogenic differentiation of DPSCs. The odontogenic differentiation genes, BMP1, ON, RUNX2, DSPP,rnhave all increased relative to the control. Our results demonstrate that C5a constitutes a positive regulator of the odontogenic DPSCrndifferentiation and identified the C5a as a key initial signal to control odontogenic DPSC differentiation.