Differential Effects of Escherichia coli– Versus Porphyromonas gingivalis–derived Lipopolysaccharides on Dental Pulp Stem Cell Differentiation in Scaffold-free Engineered Tissues

Abstract

IntroductionTo leverage the therapeutic capabilities of dental pulp stem cells (DPSCs) for regenerative endodontic applications, a better understanding of their innate defense and reparative processes is needed. Lipopolysaccharide (LPS) is a major virulent factor of gram-negative bacteria and contributor to endodontic infections. We have developed 3-dimensional scaffold-free DPSC tissues that self-organize into dentin-pulp organoids comprising a mineralized dentin-like tissue on the periphery and an unmineralized pulp-like core. In this study, scaffold-free DPSC constructs were used as controllable experimental models to study the DPSC response to bacterial challenge. MethodsScaffold-free constructs were engineered using DPSCs isolated from human third molars. To simulate bacterial exposure, DPSC constructs were exposed to either Porphyromonas gingivalis–derived LPS or Escherichia coli–derived LPS. The effects of LPS on DPSC differentiation, proliferation, and apoptosis were evaluated. ResultsEngineered tissues lacking LPS treatment self-organized into dentin-pulp organoids. LPS treatment did not negatively affect DPSC proliferation or apoptosis in the engineered tissues. Both E. coli LPS and P. gingivalis LPS inhibited the up-regulation of RUNX2 messenger RNA expression and reduced the expression of the odontoblast-associated proteins (P < .05), suggesting that LPS is inhibiting odontoblastic differentiation. However, only E. coli LPS treatment significantly reduced mineral deposition in the DPSC (P < .05) constructs, indicating that E. coli LPS but not P. gingivalis LPS reduced functional differentiation of DPSCs and prevented DPSCs from self-organizing into a dentin-pulp complex–like structure. ConclusionsThis study establishes scaffold-free DPSC constructs as models of oral disease. Furthermore, it emphasizes the diversity of LPS derived from different bacterial species and highlights the necessity of using LPS derived from clinically relevant bacteria in basic science investigations.