IADR Abstract Archives
Poloxamer-188 Alleviates Hypoxia-induced Dental Pulp Stem Cell Damage, In Vitro
Objectives: Dental caries and the resultant hypoxic conditions are reported to affect dental pulp stem cells (DPSC) and their regenerative potential in immature adult teeth. Therefore, it is beneficial to sustain and amplify DPSC to promote pulp tissue regeneration in a diseased state. It has been shown that Poloxamer 188 (Px-188) repairs damaged cell membranes by increasing lipid packing density. Hence, the objective of this study is to demonstrate the effect of Poloxamer and hypoxia on modulating the differentiation potential of human DPSC, in vitro.
Methods: DPSCs isolated from human third molar dental pulp tissue were cultured and characterized using flow cytometry analysis. DPSC (p3-p5) were cultured in a hypoxic (3%) chamber in the absence or presence of Px-188 at varying doses in odontogenic differentiation media. DPSC were then examined for their potential to differentiate and mineralize using quantitative real time PCR [qPCR] and differentiation analysis (Alizarin Red staining). Confocal microscopy was used to examine the viability of DPSC in 3-dimensional culture model using Colla CoteTM.
Results: Flow cytometry analysis revealed a procurement of 87-91% pure DPSC from human pulp tissue. DPSC grown in hypoxia for 72 hours showed a significant decrease in the potential to differentiate and mineralize. However, DPSC grown in the presence of Px-188 at concentrations 0, 1, 5 ng/ml significantly reversed its mineralization and differentiation potential. Additionally, RNA expression and immunoblot analysis of Px-188-treated DPSC also showed an upregulation of proteins essential for mineralization and differentiation. DPSC grown in 3-dimensional Colla CoteTM with Px-188 (5ng/ml) showed an increased viability when compared to cells grown in Colla CoteTM alone.
Conclusions: Px-188 treatment in hypoxic environment more effectively alleviates DPSC mineralization and differentiation potential. Therefore, our studies prove that Px-188 could potentially be used as a vehicle for drug administration to alleviate dental pulp stresses from issue inflammation.
Methods: DPSCs isolated from human third molar dental pulp tissue were cultured and characterized using flow cytometry analysis. DPSC (p3-p5) were cultured in a hypoxic (3%) chamber in the absence or presence of Px-188 at varying doses in odontogenic differentiation media. DPSC were then examined for their potential to differentiate and mineralize using quantitative real time PCR [qPCR] and differentiation analysis (Alizarin Red staining). Confocal microscopy was used to examine the viability of DPSC in 3-dimensional culture model using Colla CoteTM.
Results: Flow cytometry analysis revealed a procurement of 87-91% pure DPSC from human pulp tissue. DPSC grown in hypoxia for 72 hours showed a significant decrease in the potential to differentiate and mineralize. However, DPSC grown in the presence of Px-188 at concentrations 0, 1, 5 ng/ml significantly reversed its mineralization and differentiation potential. Additionally, RNA expression and immunoblot analysis of Px-188-treated DPSC also showed an upregulation of proteins essential for mineralization and differentiation. DPSC grown in 3-dimensional Colla CoteTM with Px-188 (5ng/ml) showed an increased viability when compared to cells grown in Colla CoteTM alone.
Conclusions: Px-188 treatment in hypoxic environment more effectively alleviates DPSC mineralization and differentiation potential. Therefore, our studies prove that Px-188 could potentially be used as a vehicle for drug administration to alleviate dental pulp stresses from issue inflammation.